Calculus bovis inhibits M2 tumor-associated macrophage polarization via Wnt/β-catenin pathway modulation to suppress liver cancer

BACKGROUND Calculus bovis(CB),used in traditional Chinese medicine,exhibits anti-tumor effects in various cancer models.It also constitutes an integral component of a compound formulation known as Pien Tze Huang,which is indicated for the treatment of liver cancer.However,its impact on the liver cancer tumor microenvironment,particularly on tumor-associated macrophages(TAMs),is not well understood.AIM To elucidate the anti-liver cancer effect of CB by inhibiting M2-TAM polarization via Wnt/β-catenin pathway modulation.METHODS This study identified the active components of CB using UPLC-Q-TOF-MS,evaluated its anti-neoplastic effects in a nude mouse model,and elucidated the underlying mechanisms via network pharmacology,transcriptomics,and molecular docking.In vitro assays were used to investigate the effects of CB-containing serum on HepG2 cells and M2-TAMs,and Wnt pathway modulation was validated by real-time reverse transcriptase-polymerase chain reaction and Western blot analysis.RESULTS This study identified 22 active components in CB,11 of which were detected in the bloodstream.Preclinical investigations have demonstrated the ability of CB to effectively inhibit liver tumor growth.An integrated approach employing network pharmacology,transcriptomics,and molecular docking implicated the Wnt signaling pathway as a target of the antineoplastic activity of CB by suppressing M2-TAM polarization.In vitro and in vivo experiments further confirmed that CB significantly hinders M2-TAM polarization and suppresses Wnt/β-catenin pathway activation.The inhibitory effect of CB on M2-TAMs was reversed when treated with the Wnt agonist SKL2001,confirming its pathway specificity.CONCLUSION This study demonstrated that CB mediates inhibition of M2-TAM polarization through the Wnt/β-catenin pathway,contributing to the suppression of liver cancer growth.

Poly(ADP-ribose)polymerase family member 14 promotes functional recovery after spinal cord injury through regulating microglia M1/M2 polarization via STAT1/6 pathway

Poly(ADP-ribose)polymerase family member 14(PARP14),which is an intracellular mono(ADP-ribosyl)transferase,has been reported to promote post-stroke functional recovery,but its role in spinal cord injury(SCI)remains unclear.To investigate this,a T10 spinal cord contusion model was established in C57BL/6 mice,and immediately after the injury PARP14 shRNA-carrying lentivirus was injected 1 mm from the injury site to silence PARP14 expression.We found that PARP14 was up-regulated in the injured spinal cord and that lentivirus-mediated downregulation of PARP14 aggravated functional impairment after injury,accompanied by obvious neuronal apoptosis,severe neuroinflammation,and slight bone loss.Furthermore,PARP14 levels were elevated in microglia after SCI,PARP14 knockdown activated microglia in the spinal cord and promoted a shift from M2-polarized microglia(anti-inflammatory phenotype)to M1-polarized microglia(pro-inflammatory phenotype)that may have been mediated by the signal transducers and activators of transcription(STAT)1/6 pathway.Next,microglia M1 and M2 polarization were induced in vitro using lipopolysaccharide/interferon-γand interleukin-4,respectively.The results showed that PARP14 knockdown promoted microglia M1 polarization,accompanied by activation of the STAT1 pathway.In addition,PARP14 overexpression made microglia more prone to M2 polarization and further activated the STAT6 pathway.In conclusion,these findings suggest that PARP14 may improve functional recovery after SCI by regulating the phenotypic transformation of microglia via the STAT1/6 pathway.

Role of N-formyl peptide receptor 2 in germinal matrix hemorrhage:an intrinsic review of a hematoma resolving pathway

Germinal matrix hemorrhage is one of the leading causes of morbidity,mortality,and acquired infantile hydrocephalus in preterm infants in the United States,with little progress made in its clinical management.Blood clots have been shown to elicit secondary brain injury after germinal matrix hemorrhage,by disrupting normal cerebrospinal fluid circulation and absorption after germinal matrix hemorrhage causing post-hemorrhagic hydrocephalus development.Current evidence suggests that rapid hematoma resolution is necessary to improve neurological outcomes after hemorrhagic stroke.Various articles have demonstrated the beneficial effects of stimulating the polarization of microglia cells into the M2 phenotype,as it has been suggested that they play an essential role in the rapid phagocytosis of the blood clot after hemorrhagic models of stroke.N-formyl peptide receptor 2(FPR2),a G-protein-coupled receptor,has been shown to be neuroprotective after stroke.FPR2 activation has been associated with the upregulation of phagocytic macrophage clearance,yet its mechanism has not been fully explored.Recent literature suggests that FPR2 may play a role in the stimulation of scavenger receptor CD36.Scavenger receptor CD36 plays a vital role in microglia phagocytic blood clot clearance after germinal matrix hemorrhage.FPR2 has been shown to phosphorylate extracellular-signal-regulated kinase 1/2(ERK1/2),which then promotes the transcription of the dual-specificity protein phosphatase 1(DUSP1)gene.In this review,we present an intrinsic outline of the main components involved in FPR2 stimulation and hematoma resolution after germinal matrix hemorrhage.

极区电离层N_(m)F_(2)观测与国际参考电离层对比研究

利用南北极极隙区与极光带纬度3个台站对电离层F2层峰值电子密度(N_(m)F_(2))长达1个太阳活动周的观测数据,对国际参考电离层IRI-2016模型在极区的适用性进行系统的定量研究。结果表明,在极光带纬度的北极Tromsø站,IRI预测与观测符合最好,大部分季节相对误差在40%以内,在太阳活动高年略好于太阳活动低年。在极隙区纬度的南极中山站和Longyearbyen站,IRI预测精度在太阳活动低年高于太阳活动高年。在中山站和北极Longyearbyen站仅个别月份相对误差在20%以内,大部分月份相对误差超过40%,冬季相对误差接近100%,特别是Longyearbyen站,在太阳活动高年冬季相对误差超过100%。从季节上看,3个台站都是冬季符合最差,夏季符合最好。IRI-2016模型对极区电离层进行预测时,难以如实反映极区等离子体对流和能量粒子沉降等极区特有的物理过程对极区电离层N_(m)F_(2)的影响。

Rutin pretreatment promotes microglial M1 to M2 phenotype polarization

Microglial cells are important resident innate immune components in the central nervous system that are often activated during neuroinflammation.Activated microglia can display one of two phenotypes,M1 or M2,which each play distinct roles in neuroinflammation.Rutin,a dietary flavonoid,exhibits protective effects against neuroinflammation.However,whether rutin is able to influence the M1/M2 polarization of microglia remains unclear.In this study,in vitro BV-2 cell models of neuroinflammation were established using 100 ng/mL lipopolysaccharide to investigate the effects of 1-hour rutin pretreatment on microglial polarization.The results revealed that rutin pretreatment reduced the expression of the proinflammatory cytokines tumor necrosis factor-α,interleukin-1β,and interleukin-6 and increased the secretion of interleukin-10.Rutin pretreatment also downregulated the expression of the M1 microglial markers CD86 and inducible nitric oxide synthase and upregulated the expression of the M2 microglial markers arginase 1 and CD206.Rutin pretreatment inhibited the expression of Toll-like receptor 4 and myeloid differentiation factor 88 and blocked the phosphorylation of I kappa B kinase and nuclear factor-kappa B.These results showed that rutin pretreatment may promote the phenotypic switch of microglia M1 to M2 by inhibiting the Toll-like receptor 4/nuclear factor-kappa B signaling pathway to alleviate lipopolysaccharide-induced neuroinflammation.

Effect of Pingchuan granule on typeⅡinnate lymphocytes and M2 polarization of macrophages in asthmatic mice

Objective:To observe the effect of Pingchuan granule on the number of typeⅡinnate lymphocytes(ILC2)and M2 polarization of macrophages in the lung tissue of asthmatic mice;Methods:Ovalbumin sensitized and challenged asthmatic mouse models were established,and then Pingchuan granules or IL-33 neutralizing antibody were given to intervene.The pathological morphology of lung tissue was observed by HE,PAS and Masson staining,and the expressions of IL-4,IL-5,IL-13 and IL-33 in BALF and lung tissue were detected by ELISA and qRT-PCR,Flow cytometry was used to detect the number of type II innate lymphocytes and type M2 macrophages in lung tissue.Western blot was used to detect the protein expression levels of ST-2,FIZZ1 and Arg-1 in lung tissue;Results:Compared with the control group,the inflammation score,PAS score and collagen staining area of the model group were significantly increased,the expressions of IL-4,IL-5,IL-13 and IL-33 in BALF and lung tissue were significantly increased,the number of ILC2 and M2 macrophages,the expression of ST-2,FIZZ1 and Arg-1 protein in lung tissue were significantly increased,and the differences were statistically significant(P<0.05);Compared with the model group,Pingchuan granule could significantly reduce the inflammation score,PAS score and collagen staining area of asthmatic mice,down-regulate the expression of IL-4,IL-5,IL-13 and IL-33 in BALF and lung tissue,reduce the number of ILC2 and M2 macrophages,and the expression of ST-2,FIZZ1 and Arg-1 protein in lung tissue(P<0.05);Conclusion:Pingchuan granule improve the airway remodeling of asthma by inhibiting the polarization of M2 macrophages mediated by ILC2.

Exosomal microRNA-588 from M2 polarized macrophages contributes to cisplatin resistance of gastric cancer cells

BACKGROUND Gastric cancer is a prevalent malignant cancer with a high incidence and significantly affects the health of modern people globally.Cisplatin(DDP)is one of the most common and effective chemotherapies for patients with gastric cancer,but DDP resistance remains a severe clinical challenge.AIM To explore the function of M2 polarized macrophages-derived exosomal microRNA(miR)-588 in the modulation of DDP resistance of gastric cancer cells.METHODS M2 polarized macrophages were isolated and identified by specific markers using flow cytometry analysis.The exosomes from M2 macrophages were identified by transmission electron microscopy and related markers.The uptake of the PKH67-labelled M2 macrophages-derived exosomes was detected in SGC7901 cells.The function and mechanism of exosomal miR-588 from M2 macrophages in the modulation of DDP resistance of gastric cancer cells was analyzed by CCK-8 assay,apoptosis analysis,colony formation assay,Western blot analysis,qPCR analysis,and luciferase reporter assay in SGC7901 and SGC7901/DDP cells,and by tumorigenicity analysis in nude mice.RESULTS M2 polarized macrophages were isolated from mouse bone marrow stimulated with interleukin(IL)-13 and IL-4.Co-cultivation of gastric cancer cells with M2 polarized macrophages promoted DDP resistance.M2 polarized macrophagesderived exosomes could transfer in gastric cancer cells to enhance DDP resistance.Exosomal miR-588 from M2 macrophages contributed to DDP resistance of gastric cancer cells.miR-588 promoted DDP-resistant gastric cancer cell growth in vivo.miR-588 was able to target cylindromatosis(CYLD)in gastric cancer cells.The depletion of CYLD reversed miR-588 inhibition-regulated cell proliferation and apoptosis of gastric cancer cells exposed to DDP.CONCLUSION In conclusion,we uncovered that exosomal miR-588 from M2 macrophages contributes to DDP resistance of gastric cancer cells by partly targeting CYLD.miR-588 may be applied as a potential therapeutic target for the treatment of gastric cancer.

Unveiling the anticancer effect of traditional Chinese herbal medicine

In this issue of World Journal of Gastroenterology,Huang et al reported that Calculus bovis(CB),a traditional Chinese herbal medicine,impedes the growth of liver cancers in vivo.Through further in vitro studies,they showed that CB suppressed the M2 polarization of tumor-associated macrophages by suppressing the Wnt signaling pathway,which consequently inhibited the growth of liver cancer.Although the effects of traditional Chinese herbal medicine are often not scientifically proven,Huang et al successfully identified the molecular mechanism involved in the anticancer effect of CB,and it is anticipated that the molecular mechanisms involved in the effects of other traditional Chinese herbal medicines will be scientifically elucidated,as demonstrated in this article.

Deployment of Polar Codes for Mission-Critical Machine-Type Communication Over Wireless Networks

Mission critical Machine-type Communication(mcMTC),also referred to as Ultra-reliable Low Latency Communication(URLLC),has become a research hotspot.It is primarily characterized by communication that provides ultra-high reliability and very low latency to concurrently transmit short commands to a massive number of connected devices.While the reduction in physical(PHY)layer overhead and improvement in channel coding techniques are pivotal in reducing latency and improving reliability,the current wireless standards dedicated to support mcMTC rely heavily on adopting the bottom layers of general-purpose wireless standards and customizing only the upper layers.The mcMTC has a significant technical impact on the design of all layers of the communication protocol stack.In this paper,an innovative bottom-up approach has been proposed for mcMTC applications through PHY layer targeted at improving the transmission reliability by implementing ultra-reliable channel coding scheme in the PHY layer of IEEE 802.11a standard bearing in mind short packet transmission system.To achieve this aim,we analyzed and compared the channel coding performance of convolutional codes(CCs),low-density parity-check(LDPC)codes,and polar codes in wireless network on the condition of short data packet transmission.The Viterbi decoding algorithm(VA),logarithmic belief propagation(Log-BP)algorithm,and cyclic redundancy check(CRC)successive cancellation list(SCL)(CRC-SCL)decoding algorithm were adopted to CC,LDPC codes,and polar codes,respectively.Consequently,a new PHY layer for mcMTC has been proposed.The reliability of the proposed approach has been validated by simulation in terms of Bit error rate(BER)and packet error rate(PER)vs.signal-to-noise ratio(SNR).The simulation results demonstrate that the reliability of IEEE 802.11a standard has been significantly improved to be at PER=10−5 or even better with the implementation of polar codes.The results also show that the general-purpose wireless networks are prominent inproviding short packet mcMTC with the modification needed.

THE VALUE OF M_2-TYPE PYRUVATE KINASE IMMUNOASSAY IN THE DIAGNOSIS OF HEPATOCARCINOMA

By using Fab'-enzyme labelled immuno absorbent assay (ELISA) with sensitivity of picogram (10-12 g or pg) level, the M-type pyruvate kinase (M-PyK) in plasma was determined in 47 cases of normal healthy adult and 26 cases of hepatocellular carcinoma (HCC) patient. It was found that the upper limits of normal male and female were 1.1 and 1.4 ng/ml (expressed as M2-PyK) respectively. The plasma M-PyK in HCC patients was significant increased to above 5 times of average normal level, the positive rate was about 95%. In 6 cases of subclinical small hepatocarcinoma and 7 cases of HCC patient with normal serum alpha-fetal protein level, the mean plasma M-PyK value was also increased. Whereas the plasma M-PyK level in acute or chronic hepatitis and other benign diseases were normal. After the HCC being resected, the plasma M-PyK returned to normal, but increased again in the cases of recurrent hepatocarcinoma, suggesting that the increased M-PyK in the plasma of HCC patient was criginated from M2-type PyK in HCC tissue. Therefore, plasma M-PyK may become a new micro-level index of hepatocarcinoma.

阶跃光纤低阶线偏振模的M^2因子分析

针对阶跃光纤输出低阶线偏振模(Linear Polarization,LP),利用二阶距定束宽,多点拟合法计算了几个低阶LPmn模相干和非相干叠加时的M2因子.结果表明,对同一LPmn模式,光纤的归一化频率参量V减小到截止频率时,M2因子出现峰值;当V远离截止频率时M2因子趋于一定值.低阶LPmn模非相干叠加时,叠加后的较高阶模成分越大,合成光束的光束质量越差,光束在一定的条件下可能比基模更接近于理想高斯光束;而相干叠加时,合成后的M2因子可以大于参与叠加的较高阶模的光束质量因子.

MCP-1 and IL-4 encapsulated hydrogel particles with macrophages enrichment and polarization capabilities for systemic lupus erythematosus treatment

Macrophages play a pivotal role in systemic lupus erythematosus(SLE)therapy.Efforts have been made to develop multifunctional drug delivery systems capable of directing macrophage polarization.Here,we present a novel hyaluronic acid methacrylate(HAMA)hydrogel microparticle encapsulating multiple cytokines for SLE remission though enhancing macrophage functions.The HAMA microparticles loaded with monocyte chemotactic protein-1(MCP-1)and interleukin-4(IL-4)were fabricated by using a microfluidic technology.The released MCP-1 facilitates the aggregation of inflammatory macrophages,after which IL-4 induces the macrophage phenotype shift from inflammatory M1 to immune-protective M2,thus restoring immune balance.We have demonstrated in MRL/lpr mice that the hydrogel microparticles could improve their efficacy of intraperitoneal drug delivery,modulate immune function,and attenuate the disease symptoms.These results suggest that our proposed microparticles delivery platform has potential clinical value for treating autoimmune diseases.

M2-type macrophage membrane-mediated delivery of Carvedilol nanocomplex for acute liver failure treatment and remodeling inflammatory microenvironment

Interactions of hepatic macrophages with local inflammatory microenvironment is the key factor promoting the development of acute liver failure(ALF).Hence,reprogramming pro-inflammatory M1 into anti-inflammatory M2 phenotype may offer a promising strategy for treating ALF by targeting inflammation.Our group found Carvedilol possessed potential anti-inflammatory property previously,which had been scarcely reported in ALF.We present a synergy strategy to induce macrophages into the phenotype M2-type anti-inflammatory macrophages with interleukin-4(IL-4)and IL-10 at first.Then Carvedilol is loaded on the macrophage membrane-camouflaged biomimetic nano-platform(termed as M2M@CNP)to evade reticuloendothelial system(RES)and afford Carvedilol delivery to the inflammatory environment with overproduced reactive oxygen species(ROS),further prolonging its circulation and accumulation.Sustainably released Carvedilol produced anti-inflammatory,antioxidant and anti-apoptosis effects,combining local M2-type cell membranes(M2-CM)inhibited pro-inflammatory cytokines and ROS levels,which in turn promoted and amplified M1 to M2 phenotype polarization efficiency.This study offers new insights into the rational design of biomimetic nanosystems for safe and effective ALF therapy to accelerate the clinical translation.

M1-type microglia can induce astrocytes to deposit chondroitin sulfate proteoglycan after spinal cord injury

After spinal cord injury(SCI),astrocytes gradually migrate to and surround the lesion,depositing chondroitin sulfate proteoglycan-rich extracellular matrix and forming astrocytic scar,which limits the spread of inflammation but hinders axon regeneration.Meanwhile,microglia gradually accumulate at the lesion border to form microglial scar and can polarize to generate a pro-inflammatory M1 phenotype or an anti-inflammatory M2 phenotype.However,the effect of microglia polarization on astrocytes is unclear.Here,we found that both microglia(CX3 CR1^(+))and astrocytes(GFAP^(+))gathered at the lesion border at 14 days post-injury(dpi).The microglia accumulated along the inner border of and in direct contact with the astrocytes.M1-type microglia(i NOS^(+)CX3 CR1^(+))were primarily observed at 3 and 7 dpi,while M2-type microglia(Arg1^(+)CX3 CR1^(+))were present at larger numbers at 7 and 14 dpi.Transforming growth factor-β1(TGFβ1)was highly expressed in M1 microglia in vitro,consistent with strong expression of TGFβ1 by microglia in vivo at 3 and 7 dpi,when they primarily exhibited an M1 phenotype.Furthermore,conditioned media from M1-type microglia induced astrocytes to secrete chondroitin sulfate proteoglycan in vitro.This effect was eliminated by knocking down sex-determining region Y-box 9(SOX9)in astrocytes and could not be reversed by treatment with TGFβ1.Taken together,our results suggest that microglia undergo M1 polarization and express high levels of TGFβ1 at 3 and 7 dpi,and that M1-type microglia induce astrocytes to deposit chondroitin sulfate proteoglycan via the TGFβ1/SOX9 pathway.The study was approved by the Institutional Animal Care and Use Committee of Anhui Medical University,China(approval No.LLSC20160052)on March 1,2016.

TMEM16F may be a new therapeutic target for Alzheimer's disease

TMEM16F is involved in many physiological processes such as blood coagulation,cell membrane fusion and bone mineralization.Activation of TMEM16F has been studied in various central nervous system diseases.High TMEM16F level has been also found to participate in microglial phagocytosis and transformation.Microglia-mediated neuroinflammation is a key factor in promoting the progression of Alzheimer’s disease.However,few studies have examined the effects of TMEM16F on neuroinflammation in Alzheimer’s disease.In this study,we established TMEM16F-knockdown AD model in vitro and in vivo to investigate the underlying regulatory mechanism about TMEM16F-mediated neuroinflammation in AD.We performed a Morris water maze test to evaluate the spatial memory ability of animals and detected markers for the microglia M1/M2 phenotype and NLRP3 inflammasome.Our results showed that TMEM16F was elevated in 9-month-old APP/PS1 mice.After TMEM16F knockdown in mice,spatial memory ability was improved,microglia polarization to the M2 phenotype was promoted,NLRP3 inflammasome activation was inhibited,cell apoptosis and Aβplaque deposition in brain tissue were reduced,and brain injury was alleviated.We used amyloid-beta(Aβ_(25-35))to stimulate human microglia to construct microglia models of Alzheimer’s disease.The levels of TMEM16F,inducible nitric oxide synthase(iNOS),proinflammatory cytokines and NLRP3 inflammasome-associated biomarkers were higher in Aβ_(25-35) treated group compared with that in the control group.TMEM16F knockdown enhanced the expression of the M2 phenotype biomarkers Arg1 and Socs3,reduced the release of proinflammatory factors interleukin-1,interleukin-6 and tumor necrosis factor-α,and inhibited NLRP3 inflammasome activation through reducing downstream proinflammatory factors interleukin-1βand interleukin-18.This inhibitory effect of TMEM16F knockdown on M1 microglia was partially reversed by the NLRP3 agonist Nigericin.Our findings suggest that TMEM16F participates in neuroinflammation in Alzheimer’s disease through participating in polarization of microglia and activation of the NLRP3 inflammasome.These results indicate that TMEM16F inhibition may be a potential therapeutic approach for Alzheimer’s disease treatment.

First principles study on lattice vibration and electrical properties of layered perovskite Sr_2M_2O_7(M=Nb, Ta)

In this paper, we performed calculations to investigate the dielectric, piezoelectric properties, Born effective charge （BEC）, and spontaneous polarization of Sr2M207, the method used in our study was a well-known density functional theory based on first-principles. The optimized results were in good agreement with previous experiments and calculations. which indicates that our calculated method is reasonable. The research we have done suggested that greater piezoelectric components of Sr2Nb207 were e31 and e33, and the contributions were derived from the A I. By studying the Born effective charge, it could be seen that the valence of ions changed, and the O of Sr2Nb207 were most obviously that caused by the covalent character of ions and the hybridization of O-2p and Nb-4d. The spontaneous polarization of Sr2Nb2O7 in the [001 ] direction is 25 p_C/cm2, while for Sr2Ta2O7, there was no spontaneous polarization in the paraelectric state. Finally, the effect of pressure on the piezoelectric properties were also investigated, the polarization of Sr2Nb2O7 decreased linearly with the increase alter pressure. All our preliminary results throw light on the nature of dielectric, piezoelectric properties, Born effective charge, and spontaneous polarization of Sr2M2OT, it was helpful for experimental research, the development of new materials, and future applications.

Are M1 and M2 macrophages Effectual Players in Pathological Conditions?

Pathologic inflammatory conditions are frequently correlated with dynamic alterations through macrophage activation,with classically activated Ml cells associated with promoting and sustaining inflammation and M2 cells implicated in resolving or smoldering chronic inflammation.Inflammation is a common feature of various chronic diseases,and it has direct involvement in the emergence and progression of these conditions.Macrophages participate in an autoregulatory loop characterizing inflammatory process,as they produce a wide range of biologically active mediators that exert either deleterious or beneficial effects during inflammation.Therefore,balancing the ratio of M1/M2 macrophages can help to ameliorate the inflammatory landscape of pathological conditions.This review will explore the role of macrophage polarization in distant pathological inflammatory conditions,such as cancer,autoimmunity,renal inflammation,stroke,and atherosclerosis,while sharing macrophage-driven pathogenesis.

Gold nanoparticle-directed autophagy intervention for antitumor immunotherapy via inhibiting tumor-associated macrophage M2 polarization

Tumor-associated macrophages(TAMs),one of the dominating constituents of tumor microenvironment,are important contributors to cancer progression and treatment resistance.Therefore,regulation of TAMs polarization from M2 phenotype towards M1 phenotype has emerged as a new strategy for tumor immunotherapy.Herein,we successfully initiated antitumor immunotherapy by inhibiting TAMs M2 polarization via autophagy intervention with polyethylene glycol-conjugated gold nanoparticles(PEG-Au NPs).PEG-Au NPs suppressed TAMs M2 polarization in both in vitro and in vivo models,elicited antitumor immunotherapy and inhibited subcutaneous tumor growth in mice.As demonstrated by the m RFP-GFP-LC3 assay and analyzing the autophagy-related proteins(LC3,beclin1 and P62),PEGAu NPs induced autophagic flux inhibition in TAMs,which is attributed to the PEG-Au NPs induced lysosome alkalization and membrane permeabilization.Besides,TAMs were prone to polarize towards M2phenotype following autophagy activation,whereas inhibition of autophagic flux could reduce the M2polarization of TAMs.Our results revealed a mechanism underlying PEG-Au NPs induced antitumor immunotherapy,where PEG-Au NPs reduce TAMs M2 polarization via induction of lysosome dysfunction and autophagic flux inhibition.This study elucidated the biological effects of nanomaterials on TAMs polarization and provided insight into harnessing the intrinsic immunomodulation capacity of nanomaterials for effective cancer treatment.

Interleukin-4 affects microglial autophagic flux

Interleukin-4 plays an important protective role in Alzheimer’s disease by regulating microglial phenotype,phagocytosis of amyloid-β,and secretion of anti-inflammatory and neurotrophic cytokines.Recently,increasing evidence has suggested that autophagy regulates innate immunity by affecting M1/M2 polarization of microglia/macrophages.However,the role of interleukin-4 in microglial autophagy is unknown.In view of this,BV2 microglia were treated with 0,10,20 or 50 ng/mL interleukin-4 for 24,48,or 72 hours.Subsequently,light chain 3-II and p62 protein expression levels were detected by western blot assay.BV2 microglia were incubated with interleukin-4(20 ng/mL,experimental group),3-methyladenine(500μM,autophagy inhibitor,negative control group),rapamycin(100 nM,autophagy inductor,positive control group),3-methyladenine+interleukin-4(rescue group),or without treatment for 24 hours,and then exposed to amyloid-β(1μM,model group)or vehicle control(control)for 24 hours.LC3-II and p62 protein expression levels were again detected by western blot assay.In addition,expression levels of multiple markers of M1 and M2 phenotype were assessed by real-time fluorescence quantitative polymerase chain reaction,while intracellular and supernatant amyloid-βprotein levels were measured by enzyme-linked immunosorbent assay.Our results showed that interleukin-4 induced microglial autophagic flux,most significantly at 20 ng/mL for 48 hours.Interleukin-4 pretreated microglia inhibited blockade of amyloid-β-induced autophagic flux,and promoted amyloid-βuptake and degradation partly through autophagic flux,but inhibited switching of amyloid-β-induced M1 phenotype independent on autophagic flux.These results indicate that interleukin-4 pretreated microglia increases uptake and degradation of amyloid-βin a process partly mediated by autophagy,which may play a protective role against Alzheimer’s disease.

Phosphatidylserine released from apoptotic cells in tumor induces M2-like macrophage polarization through the PSR-STAT3-JMJD3 axis

Background:Understanding how the tumor microenvironment is shaped by various factors is important for the development of new therapeutic strategies.Tumor cells often undergo spontaneous apoptotic cell death in tumor microen-vironment,these apoptotic cells are histologically co-localized with immunosup-pressive macrophages.However,the mechanism by which tumor cell apoptosis modulates macrophage polarization is not fully understood.In this study,we aimed to explore the tumor promoting effects of apoptotic tumor cells and the signal pathways involved.Methods:Apoptotic cells and macrophages in tumors were detected by immunohistochemical staining.Morphological analysis was performed with Giemsa staining.Lipids generated from apoptotic cells were detected by liq-uid chromatography-mass spectrometry.Phosphatidylserine-containing lipo-somes were prepared to mimic apoptotic cells.The expression of protein was determined by real-time PCR,immunohistochemistry enzyme-linked immunosorbent assay and Western blotting.Mouse malignant ascites and subcu-taneous tumor models were designed for in vivo analysis.Transgenic mice with specific genes knocked out and inhibitors specific to certain proteins were used for the mechanistic studies.Results:The location and the number of apoptotic cells were correlated with that of macrophages in several types of carcinomas.Phosphatidylserine,a lipid molecule generated in apoptotic cells,induced polarization and accumulation of M2-like macrophages in vivo and in vitro.Moreover,sustained administration of phosphoserine promoted tumor growth in the malignant ascites and subcuta-neous tumor models.Further analyses suggested that phosphoserine induced a M2-like phenotype in macrophages,which was related to the activation of phosphoserine receptors including T-cell immunoglobin mucin 4(TIM4)and the FAK-SRC-STAT3 signaling pathway as well as elevated the expression of the histone demethylase Jumonji domain-containing protein 3(JMJD3).Adminis-tration of specific inhibitors of these pathways could reduce tumor progression.Conclusions:This study suggest that apoptotic cell-generated phosphoserine might be a notable signal for immunosuppressive macrophages in tumors,and the related pathways might be potential therapeutic targets for cancer therapy.