Thoughts on the treatment of cancer pain under the guidance of traditional Chinese medicine

Chinese medicine diagnosis and treatment of cancer is a feature of traditional Chinese medicine in the prevention and treatment of major diseases.Cancer pain is one of the main clinical symptoms of cancer patients,especially those with advanced cancer,which seriously affects the quality of life of patients.At present,the principle of three-step analgesic ladder is mainly used in the treatment of cancer pain.However,the side effects such as constipation,nausea and vomiting caused by long-term use of analgesic drugs are often limited by patients'tolerance,meanwhile the pain is not effectively controlled.The mechanism of cancer pain is extremely complex.To fully understand the pathogenesis and syndromes of cancer pain in traditional Chinese,medicine is an important basis for effectively alleviating the symptoms of cancer pain.Chinese medicine has unique advantages in treating cancer pain,such as addiction,drug resistance and side effects.Based on the etiology and pathogenesis of cancer pain,this paper discusses the application of different classifications of traditional Chinese medicine in the treatment of cancer pain.

Gambogic acid induces mitochondria-dependent apoptosis by modulation of Bcl-2 and Bax in mantle cell lymphoma JeKo-1 cells

Objective： To study the mechanisms in gambogic acid （GA） -induced JeKo-1 human Mantle Cell Lymphoma cell apoptosis in vitro. Methods： The proliferation of GA-treated JeKo-1 cells was measured by CCK-8 assay and Ki-67 immunocytochemical detection. Apopt0sis, cell cycle and mitochondrial membrane potential were measured by flow cytometric analysis. Caspase-3, -8 and -9 were detected by colorimetric assay. Bcl-2 and Bax were analyzed by Western blotting. Results： GA inhibited cell growth in a time- and dose- dependent manner. GA induces apoptosis in JeKo- 1 cells but not in normal bone marrow cells, which was involved in reducing the membrane potential of mitochondria, activating caspases-3, -8 and -9 and decreasing the ratio of Bd-2 and Bax without cell cycle arresting. Conclusions： GA induced apoptosis in human MCL JeKo-1 cells by regulating Bcl-2/Bax and activating caspase-3, -8 and -9 via mitochondrial pathway without affecting cell cycle.

Initial steps on the analysis of the underlying pharmacological mechanisms of Wendan decoction on sudden deafness using network pharmacology and molecular docking

Background:Despite its widespread therapeutic use and effectiveness,the underlying pharmacologic mechanisms of Wendan decoction(WDD)and how it works to treat sudden deafness(SD)remain unclear.In this study,the pharmacological mechanisms of WDD underlying SD were analyzed using network pharmacology and molecular docking.Methods:The Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)was employed to identify the active compounds and target genes of WDD,and genes associated with SD were screened on five databases.RGUI conducted Gene Ontology(GO)functional and the Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses.A compound-target network was established using Cytoscape,and the STRING database created a protein-protein interaction(PPI)network to identify the key compounds and targets.Subsequently,a network of crucial compound-target was generated for further molecular docking analysis.For molecular docking simulations of the macromolecular target proteins and their matching ligand molecules,AutoDock Vina and AutoDockTool were utilized.Results:TCMSP identified 162 active target genes and 36 active compounds for WDD.The active target genes were compared with the 2271 genes associated with SD to identify 70 intersecting active target genes linked to 34 active compounds.The GO functional enrichment and KEGG pathway enrichment analyses were undertaken,and compound–target,and PPI networks were built.The key compounds and protein targets were identified and integrated to form a key compound–target network.Eventually,molecular docking was performed to investigate the interactions of the protein targets with their respective compounds.Conclusion:This study highlights the mechanisms of multi-compounds,targets,and pathways of WDD acting on SD and provides further evidence of crucial compounds and their matching target proteins of WDD acting on SD.

Traditional Chinese Medicine Integrated Responsive Microneedles for Systemic Sclerosis Treatment

Traditional Chinese medicine,such as Tripterygium wilfordii and Paeonia lactiflora,has potential values in treating systemic sclerosis(SSc)and other autoimmune diseases,while their toxic side effect elimination and precise tropical drug delivery are still challenges.Here,we present multiple traditional Chinese medicine integrated photoresponsive black phosphorus(BP)microneedles(MNs)with the desired features for the SSc treatment.By employing a template-assisted layer-by-layer curing method,such MNs with triptolide(TP)/paeoniflorin(Pae)needle tips and BP-hydrogel needle bottoms could be well generated.The combined administration of TP and Pae can not only provide anti-inflammatory,detoxification,and immunomodulatory effects to treat skin lesions in the early stage of SSc but also remarkably reduce the toxicity of single drug delivery.Besides,the additive BPs possess good biocompatibility and near-infrared(NIR)responsiveness,imparting the MN photothermal-controlled drug release capability.Based on these features,we have demonstrated that the traditional Chinese medicine integrated responsive MNs could effectively improve skin fibrosis and telangiectasia,reduce collagen deposition,and reduce epidermal thickness in the SSc mouse models.These results indicated that the proposed Chinese medicine integrated responsive MNs had enormous potential in clinical therapy of SSc and other diseases.

Chemical Constituents and Pharmacological Properties of Frankincense:Implications for Anticancer Therapy

The discovery of novel antitumor agents derived from natural plants is a principal objective of anticancer drug research.Frankincense,a widely recognized natural antitumor medicine,has undergone a systematic review encompassing its species,chemical constituents,and diverse pharmacological activities and mechanisms.The different species of frankincense include Boswellia serrata,Somali frankincense,Boswellia frereana,and Boswellia arabica.Various frankincense extracts and compounds exhibit antitumor,anti-inflammatory,and hepatoprotective properties and antioxidation,memory enhancement,and immunological regulation capabilities.They also have comprehensive effects on regulating flora.Frankincense and its principal chemical constituents have demonstrated promising chemoprophylactic and therapeutic abilities against tumors.This review provides a systematic summary of the mechanism of action underlying the antitumor effects of frankincense and its major constituents,thus laying the foundations for developing effective tumor-combating targets.

Construction of potential idiopathic pulmonary fibrosis related microRNA and messenger RNA regulatory network

To the Editor:Idiopathic pulmonary fibrosis(IPF)is a kind of lung disease characterized by chronic and progressive pulmonary fibrosis with unknown etiology,whose pathological manifestation is usual interstitial pneumonia.111 However,the prognosis of IPF was still poor,and the median survival time after diagnosis was about 2 to 3 years Therefore,it seems important to seek and develop an effective therapeutic modality for IPF.Through base pairs of intramolecular complementary sequences of messenger RNA(mRNA),microRNA(miRNA)plays a key role in RNA silencing and post-transcriptional gene expression regulation.

MFG-E8 Alleviates Cognitive Impairments Induced by Chronic Cerebral Hypoperfusion by Phagocytosing Myelin Debris and Promoting Remyelination

Chronic cerebral hypoperfusion is one of the pathophysiological mechanisms contributing to cognitive decline by causing white matter injury.Microglia phagocytosing myelin debris in a timely manner can promote remyelination and contribute to the repair of white matter.However,milk fat globule-epidermal growth factor-factor 8(MFG-E8),a microglial phagocytosis-related protein,has not been well studied in hypoperfusion-related cognitive dysfunction.We found that the expression of MFG-E8 was significantly decreased in the brain of mice after bilateral carotid artery stenosis(BCAS).MFG-E8 knockout mice demonstrated more severe BCAS-induced cognitive impairments in the behavioral tests.In addition,we discovered that the deletion of MFG-E8 aggravated white matter damage and the destruction of myelin microstructure through fluorescent staining and electron microscopy.Meanwhile,MFG-E8 overexpression by AAV improved white matter injury and increased the number of mature oligodendrocytes after BCAS.Moreover,in vitro and in vivo experiments showed that MFG-E8 could enhance the phagocytic function of microglia via theαVβ3/αVβ5/Rac1 pathway and IGF-1 production to promote the differentiation of oligodendrocyte progenitor cells into mature oligodendrocytes.Interestingly,we found that MFG-E8 was mainly derived from astrocytes,not microglia.Our findings suggest that MFG-E8 is a potential therapeutic target for cognitive impairments following cerebral hypoperfusion.

Jug-PLGA-NPs, a New Form of Juglone with Enhanced Efficiency and Reduced Toxicity on Melanoma

Objective: To verrify the anti-tumor efficacy and toxicity between juglone(Jug) and Jug-loaded poly lactic-co-glycolic acid(PLGA) nanoparticles(Jug-PLGA-NPs). Methods: Jug-PLGA-NPs were prepared by ultrasonic emulsification. The anti-tumor activity of Jug(2, 3, 4 μg/mL) and Jug-PLGA-NPs(Jug: 2, 3, 4 μg/mL) in vitro was measured by MTT assay and cell apoptosis analysis. The distribution, anti-tumor effect and biological safety in vivo was evaluated on A375 nude mice. Results: With the advantage of good penetration and targeting properties, Jug-PLGA-NPs significantly inhibited proliferation and migration of melanoma cells both in vitro and in vivo(P<0.05 or P<0.01) with acceptable biocompatibility. Conclusions: Jug can inhibit the growth of melanoma but is highly toxic. With the advantage of sustained release, tumor targeting, anti-tumor activity and acceptable biological safety, Jug-PLGA-NPs provide a new pharmaceutical form for future application of Jug.

First-line penpulimab(an anti-PD1 antibody)and anlotinib(an angiogenesis inhibitor)with nab-paclitaxel/gemcitabine(PAAG)in metastatic pancreatic cancer:a prospective,multicentre,biomolecular exploratory,phase II trial

Metastatic pancreatic cancer(mPC)has a dismal prognosis.Herein,we conducted a prospective,multicentre,single-arm,phase II trial evaluating the efficacy and safety of penpulimab and anlotinib in combination with nab-paclitaxel/gemcitabine(PAAG)in patients with first-line mPC(NCT05493995).The primary endpoints included the objective response rate(ORR)and disease control rate(DCR),while secondary endpoints encompassed progression-free survival(PFS),overall survival(OS),and safety.In 66 patients analysed for efficacy,the best response,indicated by the ORR,was recorded at 50.0%(33/66)(95%CI,37.4–62.6%),with 33 patients achieving partial response(PR).Notably,the DCR was 95.5%(63/66,95%CI,87.3–99.1%).The median PFS(mPFS)and OS(mOS)were 8.8(95%CI,8.1–11.6),and 13.7(95%CI,12.4 to not reached)months,respectively.Grade 3/4 treatment-related adverse events(TRAEs)were reported in 39.4%of patients(26/66).In prespecified exploratory analysis,patients with altered SWI/SNF complex had a poorer PFS.Additionally,low serum CA724 level,high T-cell recruitment,low Th17 cell recruitment,and high NK CD56dim cell scores at baseline were potential predicative biomarkers for more favourable efficacy.In conclusion,PAAG as a first-line therapy demonstrated tolerability with promising clinical efficacy for mPC.The biomolecular findings identified in this study possess the potential to guide the precise clinical application of the triple-combo regimen.

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.

Enhanced therapeutic effects of apoptotic cell-conditioned mesenchymal stem cells in lupus-prone MRL/lpr mice

Background::Apoptotic cell-conditioned mesenchymal stem cells (AC-MSCs) exhibit stronger T cell suppressive ability via cyclooxygenase 2 (COX2)/prostaglandin E2 (PGE2);however, whether AC-MSCs exhibit enhanced therapeutic effects on systemic lupus erythematosus (SLE) remains unknown. Methods: Splenocytes from MRL/MPJ-Fas lpr (MRL /lpr) mice were cocultured with AC-MSCs, and the proportion of plasma cells was determined by flow cytometry. MSCs, AC-MSCs, COX2 knockdown MSCs, and COX2 knockdown AC-MSCs were infused into MRL/ lpr mice ( n = 10/group). Survival rates and lupus symptoms, including proteinuria, kidney injury, renal immune complex deposition, and autoantibody production, were assessed. Additionally, the number of plasma cells and serum levels of inflammatory cytokines were measured. Results::The AC-MSCs significantly inhibited plasma cells via PGE2 after 24 h coculture in vitro, whereas MSCs did not. In the MRL /lpr mice, AC-MSC treatment led to a significantly higher survival rate than phosphate-buffered saline (PBS) treatment (90% vs. 50%, p < 0.05). Moreover, AC-MSC infusion decreased urine protein levels as early as 1 week after administration (0.89 ± 0.55 mg/mL vs. 1.59 ± 0.60 mg/mL, p < 0.05, compared with PBS treatment). Administration of both MSCs and AC-MSCs reduced renal immunoglobulin G and complement C3 deposition, whereas COX2 knockdown MSCs and COX2 knockdown AC-MSCs did not. Serum anti-dsDNA antibody levels in AC-MSC-treated mice significantly decreased (0.40 ± 0.25 vs. 0.99 ± 0.58, p < 0.05), compared with PBS treatment, as well as the number of plasma cells in both the spleen ([2.14 ± 1.05] × 10^(6) vs. [8.02 ± 4.01] × 10^(6), p < 0.01) and renal-draining lymph nodes ([0.78 ± 0.68] × 10^(6) vs. [2.49 ± 1.45] × 10^(6), p < 0.05). Additionally, AC-MSCs inhibited the production of inflammatory cytokines, including interleukin-21, tumor necrosis factor-alpha, and monocyte chemoattractant protein-1. Conclusions::AC-MSCs enhanced the therapeutic effects in mice with lupus, which were partially mediated by COX2/PGE2. Therefore, AC preconditioning may be a new strategy for MSC transplantation in the treatment of SLE.

Transplantation of Mesenchymal Stem Cells Attenuates Acute Liver Failure in Mice via an Interleukin-4-dependent Switch to the M2 Macrophage Anti-inflammatory Phenotype

Background and Aims:Transplantation of mesenchymal stem cells(MSCs)derived from bone marrow(BM)is an alternative treatment of acute liver failure(ALF)mainly be-cause of the resulting anti-inflammatory activity.It is not known how MSCs regulate local immune responses and liver regeneration.This study explored the effects of MSCs on hepatic macrophages and the Wnt signaling pathway in ALF.Methods:MSCs were isolated from BM aspirates of C57BL/6J mice,and transplanted in mice with ALF induced by D-galactosamine(D-Gal).The proliferation of hepato-cytes was assayed by immunohistochemical(IHC)staining of Ki-67 and proliferating cell nuclear antigen(PCNA).The levels of key proteins in the Wnt signaling pathway were assayed by western blotting and cytokines were determined enzyme-linked immunosorbent assays(ELISAs).A mac-rophage polarization assay characterized the M1/M2 ratio.The potential role of interleukin-4(IL-4)in the biological ac-tivity of MSCs was determined by silencing of IL-4.Results:Transplantation of allogeneic MSCs significantly attenuated D-Gal-induced hepatic inflammation and promoted liver re-generation.MSC transplantation significantly promoted a phenotypic switch from proinflamatory M1 macrophages to anti-inflammatory M2 macrophages,leading to significant Wnt-3a induction and activation of the Wnt signaling path-way in mice with D-Gal-induced ALF.Of the paracrine fac-tors secreted by MSCs(G-CSF,IL-6,IL-1 beta,IL-4,and IL-17A),IL-4 was specifically induced following transplantation in the ALF model mice.The silencing of IL-4 significantly ab-rogated the phenotypic switch to M2 macrophages and the protective effects of MSCs in both the ALF model mice and a co-culture model in an IL-4 dependent manner.Conclu-sions:In vivo and in vitro studies showed that MSCs ame-liorated ALF through an IL-4-dependent macrophage switch toward the M2 anti-inflammatory phenotype.The findings may have clinical implications in that overexpression of IL-4 may enhance the therapeutic effects of allogeneic MSC transplantation in the treatment of ALF.

Puerarin improves diabetic wound healing via regulation of macrophage M2 polarization phenotype

Background:Skin wound healing depends on the progress of different but overlapping stages of healing,including hemostasis,inflammatory,proliferative and remodeling.Failure of these stages to occur in a timely and gradual manner may result in non-healing pathological wounds.Macrophages and neutrophils have been shown to play an essential role in the inflammatory responses of wound tissue,and their active plasticity allows them to modulate tissue damage and repair functions.The ability of macrophages and neutrophils to regulate the occurrence and resolution of inflammatory processes is essential for the treatment of pathological wound healing.Methods:Mice were categorized into negative control,streptozotocin,streptozotocin+puerarin and puerarin groups.The traditional Chinese medicine extract puerarin was selected to treat different groups of mice with a full-thickness skin defect wound.Cells of the RAW264.7 cell line were stimulated under different puerarin conditions.Then,real time quantitative polymerase chain reaction(RT-qPCR),western blot,immunofluorescence and other assays were carried out to explore the effect of puerarin on wound healing and its molecular mechanism.Results:Animal experiments found that the wound healing of diabetic mice treated with puerarin was significantly accelerated,and histological analysis found that puerarin treatment markedly decreased the infiltration of macrophages and neutrophils in wound tissue.Through western blot,RT-qPCR and immunofluorescence experiments,it was observed that puerarin treatment remarkably inhibited nuclear factor kinase B(NF-κB)and mitogen-activated protein kinase(MAPK)signaling pathways,downregulated the expression of inflammatory cytokines and induced the M2 polarization of macrophages.At the cellular level,we also observed that puerarin improved M2 macrophage polarization and inhibited inflammatory pathway activation in a high-glucose culture.Conclusion:Puerarin has a significant therapeutic effect on wound healing in diabetic mice.The therapeutic effect is achieved by regulating macrophage polarization through suppressing NF-κB and MAPK signaling cascades.

Neuronal NR4A1 deficiency drives complement-coordinated synaptic stripping by microglia in a mouse model of lupus

Neuropsychiatric lupus(NPSLE)is a frequent manifestation of systemic lupus erythematosus(SLE)that occurs in 40-90%of SLE patients;however,the underlying mechanisms remain elusive,causing a severe lack of therapeutic targets for this condition.Here,we show that complement-coordinated elimination of synapses participated in NPSLE in MRL/lpr mice,a lupus-prone murine model.We demonstrated that lupus mice developed increased anxiety-like behaviors and persistent phagocytic microglial reactivation before overt peripheral lupus pathology.

Renaming NAFLD to MAFLD:Advantages and Potential Changes in Diagnosis,Pathophysiology,Treatment,and Management

In recent years,with the increasing incidence of obesity and other metabolic diseases,the prevalence of non-alcoholic fatty liver disease(NAFLD)has increased and it has become a major health problem affecting more than one quarter of the world’s population.Recently,experts reached a consensus that NAFLD does not reflect the current knowledge,and metabolic dysfunctionassociated fatty liver disease(MAFLD)was suggested as a more appropriate term.MAFLD is not just a simple renaming of NAFLD.The definition of MAFLD allows a patient to have dual(or more)etiologies for their liver disease,which will help to exclude more heterogeneous patients.In this review,we introduce the significant differences between the definitions of NAFLD and MAFLD.In addition,we also describe the advantages of the term MAFLD in the pathophysiology,therapy,and patient management.

Compound from Magnolia officinalis Ameliorates White Matter Injury by Promoting Oligodendrocyte Maturation in Chronic Cerebral Ischemia Models

Chronic cerebral hypoperfusion leads to white matter injury(WMI),which subsequently causes neurodegeneration and even cognitive impairment.However,due to the lack of treatment specifically for WMI,novel recognized and effective therapeutic strategies are urgently needed.In this study,we found that honokiol and magnolol,two compounds derived from Magnolia officinalis,significantly facilitated the differentiation of primary oligodendrocyte precursor cells(OPCs)into mature oligodendrocytes,with a more prominent effect of the former compound.Moreover,our results demonstrated that honokiol treatment improved myelin injury,induced mature oligodendrocyte protein expression,attenuated cognitive decline,promoted oligodendrocyte regeneration,and inhibited astrocytic activation in the bilateral carotid artery stenosis model.Mechanistically,honokiol increased the phosphorylation of serine/threonine kinase(Akt)and mammalian target of rapamycin(mTOR)by activating cannabinoid receptor 1 during OPC differentiation.Collectively,our study indicates that honokiol might serve as a potential treatment for WMI in chronic cerebral ischemia.

Activation of NLRP3 Inflammasome via Drp1 Overexpression in Kupffer Cells Aggravates Ischemia-reperfusion Injury in Hepatic Steatosis

Background and Aims:Donors with fatty livers are considered to address the shortage of livers for transplantation,but those livers are particularly sensitive to ischemia-reperfusion injury(IRI),and an increased incidence of graft failure is observed.Kupffer cells account for 20–35%of liver nonparenchymal cells,and have been shown to participate in the process of IRI and inflammatory reactions of hepatic steatosis.NOD-like receptor thermal protein domain-associated protein 3(NLRP3)is an intracellular sensor activated by Kupffer cells to promote generation and participates in IRI.Dynamics-associated protein 1(Drp1)is one of the main proteins regulating mitochondrial division and exacerbates IRI by affecting mitochondrial dynamics.The mechanism of interaction of Kupffer cells with Drp1 and NLRP3 to aggravate IRI has not been clarified.Methods:A mouse model of hepatic steatosis was established by feeding the mice with a high-fat diet.In vitro experiments were performed using AML12 normal mouse liver cells and RAW264.7 mononuclear macrophage cells cultured in medium with palmitate and oleic acid.Western blotting and immunohistochemical(IHC)staining were used to detect the expression of NLRPP3 and Drp1 in IRI in the control and high-fat diet groups.The expression of F4/80+cells during IRI in hepatic steatosis was verified by IHC staining,and the role of NLRPP3 and Drp1 in Kupffer-cell mediated IRI was investigated by targeting Drp-1 inhibition.Results:Drp1 and NLRP3 expression was increased during IRI in hepatic steatosis,and the expression of Drp1 and NLRP3 were decreased after the elimination of Kupffer cells.That indicated Kupffer cells were involved in the process of IRI in hepatic steatosis through the action of Drp1 and NLRP3.After Drp1 inhibition,liver function was restored and NLRP3 expression level was reduced.Conclusions:Kupffer cells aggravated IRI in hepatic steatosis via NLRP3 and Drp1.Drp1 inhibitors might be useful as specific therapeutics to alleviate IRI in hepatic steatosis and may have promise in case of liver donor shortage.

Treatment of Alzheimer’s disease by microcapsule regulates neurotransmitter release via microfluidic technology

Alzheimer’s disease(AD)is a progressive neurodegenerative disease with a complex etiology.The main neu-ropathological feature is the accumulation of amyloid-beta(Aβ),and the dysregulation of the cholinergic system is well associated with its mechanism of occurrence,for which no effective treatment is yet available.Daily oral administration remains the mainstay of treatment with AD,and how to improve the efficacy,prolong adsorp-tion and medication compliance is still the focus of the current solution.We proposed a microcapsule based on microfluidic electrospray to form an intestinal epithelial lining for AD treatment,reducing the frequency of administration.Microfluidic electrospray technology was recruited to overcome the limitations associated with the variability in the microencapsulation production process and to produce functional microcapsules with finely adapted chemical composition,capsule thickness and encapsulant volume ratio.These microcapsules could slowly release drugs after adhering to the intestine,and their effectiveness and safety were further evaluated using cell culture studies and animal model studies.The results from the in vivo and in vitro experiments showed a significant reduction in administration frequency(i.e.,from daily medication to once every five days),superior therapeutic efficacy and sufficient safety of these microcapsules in cell culture and APP/PS1 mice.These features make the microcapsules an excellent drug delivery system and represent great potential for clinical applications in AD.

心肌缺血再灌注损伤中心肌免疫细胞单细胞图谱绘制及具有心肌保护效应的Ym-1hi中性粒细胞的鉴定

Myocardial ischemia–reperfusion injury(MIRI)is a major hindrance to the success of cardiac reperfusion therapy.Although increased neutrophil infiltration is a hallmark of MIRI,the subtypes and alterations of neutrophils in this process remain unclear.Here,we performed single-cell sequencing of cardiac CD45^(+)cells isolated from the murine myocardium subjected to MIRI at six-time points.We identified diverse types of infiltrating immune cells and their dynamic changes during MIRI.Cardiac neutrophils showed the most immediate response and largest changes and featured with functionally heterogeneous subpopulations,including Ccl3^(hi)Neu and Ym-1^(hi)Neu,which were increased at 6 h and 1 d after reperfusion,respectively.Ym-1^(hi)Neu selectively expressed genes with protective effects and was,therefore,identified as a novel specific type of cardiac cell in the injured heart.Further analysis indicated that neutrophils and their subtypes orchestrated subsequent immune responses in the cardiac tissues,especially instructing the response of macrophages.The abundance of Ym-1^(hi)Neu was closely correlated with the therapeutic efficacy of MIRI when neutrophils were specifically targeted by anti-Lymphocyte antigen 6 complex locus G6D(Ly6G)or anti-Intercellular cell adhesion molecule-1(ICAM-1)neutralizing antibodies.In addition,a neutrophil subtype with the same phenotype as Ym-1^(hi)Neu was detected in clinical samples and correlated with prognosis.Ym-1 inhibition exacerbated myocardial injury,whereas Ym-1 supplementation significantly ameliorated injury in MIRI mice,which was attributed to the tilt of Ym-1 on the polarization of macrophages toward the repair phenotype in myocardial tissue.Overall,our findings reveal the antiinflammatory phenotype of Ym-1^(hi)Neu and highlight its critical role in myocardial protection during the early stages of MIRI.

Cortical Organoid-on-a-Chip with Physiological Hypoxia for Investigating Tanshinone IIA-Induced Neural Differentiation

Cortical organoids represent cutting-edge models for mimic human brain development during the early and even middle stage of pregnancy, while they often fail to recreate the complex microenvironmental factors, such as physiological hypoxia. Herein, to recapitulate fetal brain development, we propose a novel cortical organoid-on-a-chip with physiological hypoxia and further explore the effects of tanshinone IIA (Tan IIA) in neural differentiation. The microfluidic chip was designed with a micropillar array for the controlled and efficient generation of cortical organoids. With low oxygen, the generated cortical organoids could recapitulate key aspects of early-gestational human brain development. Compared to organoids in normoxic culturing condition, the promoted neurogenesis, synaptogenesis and neuronal maturation were observed in the present microsystem, suggesting the significance of physiological hypoxia in cortical development. Based on this model, we have found that Chinese herbal drug Tan IIA could promote neural differentiation and maturation, indicating its potential therapeutic effects on neurodevelopmental disorders as well as congenital neuropsychiatric diseases. These results indicate that the proposed biomimetic cortical organoid-on-a-chip model with physiological hypoxia can offer a promising platform to simulate prenatal environment, explore brain development, and screen natural neuroactive components.