Mechanistic engineering of celastrol liposomes induces ferroptosis and apoptosis by directly targeting VDAC2 in hepatocellular carcinoma

Hepatocellular carcinoma(HCC)is one of most common and deadliest malignancies.Celastrol(Cel),a natural product derived from the Tripterygium wilfordii plant,has been extensively researched for its potential effectiveness in fighting cancer.However,its clinical application has been hindered by the unclear mechanism of action.Here,we used chemical proteomics to identify the direct targets of Cel and enhanced its targetability and antitumor capacity by developing a Cel-based liposomes in HCC.We demonstrated that Cel selectively targets the voltage-dependent anion channel 2(VDAC2).Cel directly binds to the cysteine residues of VDAC2,and induces cytochrome C release via dysregulating VDAC2-mediated mitochondrial permeability transition pore(mPTP)function.We further found that Cel induces ROS-mediated ferroptosis and apoptosis in HCC cells.Moreover,coencapsulation of Cel into alkyl glucoside-modified liposomes(AGCL)improved its antitumor efficacy and minimized its side effects.AGCL has been shown to effectively suppress the proliferation of tumor cells.In a xenograft nude mice experiment,AGCL significantly inhibited tumor growth and promoted apoptosis.Our findings reveal that Cel directly targets VDAC2 to induce mitochondria-dependent cell death,while the Cel liposomes enhance its targetability and reduces side effects.Overall,Cel shows promise as a therapeutic agent for HCC.

领导信任对幼儿园教师工作满意度的影响:角色超载的中介作用

本研究采用问卷调查法,基于两个时间点286名幼儿园教师的匹配数据探讨了领导信任对幼儿园教师工作满意度的影响机制,并运用描述性统计分析、相关分析和层次回归分析等方法进行验证,结果发现领导信任与幼儿园教师的工作满意度呈显著负向关系;领导信任与幼儿园教师的角色超载呈显著正向关系;角色超载与幼儿园教师的工作满意度呈显著负向关系;角色超载在领导信任对幼儿园教师工作满意度的影响中起中介作用;领导信任对持有高互惠信念的幼儿园教师的角色超载具有更强的正向影响。这表明"信任未必多多益善",幼儿园领导者应正确理解信任可能产生的负面影响,密切关注教师的情绪变化,缓解教师的工作压力;针对不同互惠信念的教师采取不同的管理方式,培养教师团队的合作意识;通过合理授权增强教师的认同感,主动同教师交流和沟通,由此提高教师的工作满意度。

挑战性—阻碍性压力对幼儿园教师自我效能的影响:有调节的中介模型

以往关于幼儿园教师工作压力的研究集中于各种压力对个体的影响,忽视了对压力属性的区分。本研究从挑战性压力与阻碍性压力的区别出发,通过问卷法对312名幼儿园教师进行调查,探究教师所受挑战性压力与阻碍性压力的基本状况及其与教师自我效能、情绪耗竭、认知重评之间的关系。结果表明,幼儿园教师所受挑战性压力与阻碍性压力总体水平较高,不同学历与有无编制的教师在挑战性压力上有显著差异;不同教龄与不同等级幼儿园的教师在两种压力上都存在显著差异;挑战性压力、阻碍性压力均负向预测自我效能;情绪耗竭在挑战性压力对自我效能的影响中起完全中介作用,而在阻碍性压力对自我效能的影响中不起中介作用;认知重评在两种压力与情绪耗竭之间起调节作用。有效疏解幼儿园教师工作压力,需要多种措施相互配合。首先,幼儿园领导应合理控制挑战性压力,积极削减阻碍性压力,尤其要关注本科及以上学历、有编制、教龄在20年以上的教师所受挑战性压力的水平。其次,应针对认知重评水平不同的教师,采取差异化的管理措施,培养教师积极乐观应对压力的心态。再次,应给予教师充分的物质层面的支持和精神情感层面的关怀,强化其自我效能感和职业幸福感。

Regulation of Hepatitis C Virus Replication and Gene Expression by the MAPK-ERK Pathway

The mitogen activated protein kinases-extracellular signal regulated kinases （MAPK-ERK） pathway is involved in regulation of multiple cellular processes including the cell cycle. In the present study using a Huh7 cell line Conl with an HCV replicon, we have shown that the MAPK-ERK pathway plays a significant role in the modulation of HCV replication and protein expression and might influence IFN-a signalling. Epithelial growth factor （EGF） was able to stimulate ERK activation and decreased HCV RNA load while a MAPK-ERK pathway inhibitor U0126 led to an elevated HCV RNA load and higher NS5A protein amounts in Conl cells. It could be further demonstrated that the inhibition of the MAPK-ERK pathway facilitated the translation directed by the HCV internal ribosome entry site. Consistently, a U0126 treatment enhanced activity of the HCV reporter replicon in transient transfeetion assays. Thus, the MAPK-ERK pathway plays an important role in the regulation of HCV gene expression and replication. In addition, cyclin-dependent kinases （CDKs） downstream of ERK may also be involved in the modulation of HCV replication since roscovitine, an inhibitor of CDKs had a similar effect to that of U0126. Modulation of the cell cycle progression by cell cycle inhibitor or RNAi resulted consistently in changes of HCV RNA levels. Further, the replication of HCV replicon in Conl cells was inhibited by IFN-~z. The inhibitory effect of IFN-CZ could be partly reversed by pre-incubation of Con-1 cells with inhibitors of the MAPK-ERK pathway and CDKs. It could be shown that the MAPK-ERK inhibitors are able to partially modulate the expression of interferon-stimulated genes.

多维异质异构大型构件智能增材制造研究进展

电弧增材是近年发展起来的一种高效率、低成本、高性能、低精度整体制造方法,可成形超高强钢、轻合金等多种金属构成的一体化高性能构件.电弧-激光复合、增材-形变、增材-减材等复合成形技术,可进一步提高成形精度,提升构件韧性,更好地成形异质异构构件.本文从多维异质异构概念内涵、电弧复合增材技术、电弧增材过程智能控制等方面对多维异质异构大型构件智能电弧增材技术进行了综述,重点分析了增材过程参数-熔池视觉-应力-变形等协同传感技术;利用深度学习等人工智能方法,在线调整工艺参数,控制缺陷、抑制应力、减小变形,研制的大型多维异质构件多机器人智能复合增材装备,最大可增材10 m×4 m×4 m多金属构件;分析了电弧增材构件微观组织演变、静(动)态力学性能和抗超高速冲击性能特征;最后,指出了多维异质异构增材技术的4大发展趋势.

Phylogenetic diversity and bioactivity of culturable deepsea-derived fungi from Okinawa Trough

Deep-sea sediments are now recognized as a home for rich and largely microbial community.Recently,it has been believed in an increasing number of studies that bacteria could be abundant in deepsea sediments of many types;however,fungi in deep-sea sediments remain relatively unknown.The phylogenetic diversity and bioactivity of culturable deep-sea-derived fungi from Okinawa Trough sediments were investigated in traditional method combined with fungal identification of molecular biology in this study.A total of 76 isolates belonged to 15 fungal taxa were recovered in a harsh condition of low nutrient and low temperature,indicating that the fungal communities in deep-sea sediments from Okinawa Trough were relatively abundant and diversified.Aspergillus,Cladosporium,and Penicillium were the dominant fungal genera,while Mycosphaerella,Purpureocillium,and Schizophyllum were relatively rare in the deep-sea sediments from Okinawa Trough.Among the six genera recovered,Mycosphaerella was firstly recovered from deep-sea sediments in this study.Moreover,about 75%of the extracts from the 15 fungal representative isolates displayed distinct bioactivity against at least one indicator bacterium or marine macrofouler,emphasizing the potentials of these deep-sea-derived fungi from Okinawa Trough as producers of bioactive metabolites.Notably,isolates Cladosporium oxysporum SCSIO z001 and Penicillium citrinum SCSIO z049 displayed a wide spectrum of bioactivities,isolates Cladosporium cladosporioides SCSIO z015,Cladosporium sphaerospermum SCSIO z030,and Penicillium verruculosum SCSIO z007 exhibited a strong anti-bacterial-growth activity,and isolate Penicillium chrysogenum SCSIO z062 displayed a strong anti-larval-settlement activity.These results suggest that these isolates deserved further study as potential sources of novel bioactive metabolites.

Molecular cloning and structural analysis of human norepinephrine transporter gene(NETHG)

A cDNA molecule encoding a major part of the hu-man Norepinephrine transporter(hNET) was synthesized by means of Polymerase Chain Reaction(PCR) technique and used as a probe for selecting the human genomic NET gene. A positive clone harbouring the whole gene was ob-tained from a human lymphocyte genomic library through utilizing the "genomic walking" technique. The clone, des-ignated as phNET, harbours a DNA fragment of about 59 kb in length inserted into BamH Ⅰ site in cosmid pWE15.The genomic clone contains 14 exons encoding all amino acid residues in the protein. A single exon encodes a dis-tinct transmembrane domaill, except for transmembrane domain 10 and 11, which are encoded by part of two ex-ons respectively, and exon 12, which encodes part of do-main 11 and all of domain 12. These results imply that there is a close relationship between exon splicing of a gene and structural domains of the protein, as is the case for the human γ-aminobutyric acid transporter(hGAT) and a number of other membrane proteins.

A kinetic resolution accompanied chirality transformation process for asymmetric synthesis of chiral phenol,oxaziridine,andα-hydroxyl cyclic ketone

Chirality transformation is a basic,attractive,and important strategy for obtaining enantioenriched products with desired chiral elements.The reported chirality conversion reaction often involves the process from one type of chirality to another one.To better utilize the chirality transformation strategy for obtaining two or more products with different chiral elements in a single reaction,a new method of kinetic resolution accompanied by a chirality transformation protocol is proposed and successfully realized in this study.This process is used for the asymmetric oxidation of phenol compounds along with the kinetic resolution of oxaziridines.A wide scope of products,including axially chiral phenols,oxaziridines,andα-hydroxyl cyclic ketones were smoothly obtained in high levels of yields and enantioselectivities in the developed method.These products can be readily used for the synthesis of various types of chiral ligands,which are potential choices for other catalytic asymmetric reactions.

Elite Dung Beetle Optimization Algorithm for Multi-UAV Cooperative Search in Mountainous Environments

This paper aims to address the problem of multi-UAV cooperative search for multiple targets in a mountainous environment,considering the constraints of UAV dynamics and prior environmental information.Firstly,using the target probability distribution map,two strategies of information fusion and information diffusion are employed to solve the problem of environmental information inconsistency caused by different UAVs searching different areas,thereby improving the coordination of UAV groups.Secondly,the task region is decomposed into several high-value sub-regions by using data clustering method.Based on this,a hierarchical search strategy is proposed,which allows precise or rough search in different probability areas by adjusting the altitude of the aircraft,thereby improving the search efficiency.Third,the Elite Dung Beetle Optimization Algorithm(EDBOA)is proposed based on bionics by accurately simulating the social behavior of dung beetles to plan paths that satisfy the UAV dynamics constraints and adapt to the mountainous terrain,where the mountain is considered as an obstacle to be avoided.Finally,the objective function for path optimization is formulated by considering factors such as coverage within the task region,smoothness of the search path,and path length.The effectiveness and superiority of the proposed schemes are verified by the simulation.

A novel engineered IL-21 receptor arms T-cell receptor-engineered T cells(TCR-T cells)against hepatocellular carcinoma

Strategies to improve T cell therapy efficacy in solid tumors such as hepatocellular carcinoma(HCC)are urgently needed.The common cytokine receptorγchain(γc)family cytokines such as IL-2,IL-7,IL-15 and IL-21 play fundamental roles in T cell development,differentiation and effector phases.This study aims to determine the combination effects of IL-21 in T cell therapy against HCC and investigate optimized strategies to utilize the effect of IL-21 signal in T cell therapy.The antitumor function of AFP-specific T cell receptor-engineered T cells(TCR-T)was augmented by exogenous IL-21 in vitro and in vivo.IL-21 enhanced proliferation capacity,promoted memory differentiation,downregulated PD-1 expression and alleviated apoptosis in TCR-T after activation.A novel engineered IL-21 receptor was established,and TCR-T armed with the novel engineered IL-21 receptors(IL-21R-TCR-T)showed upregulated phosphorylated STAT3 expression without exogenous IL-21 ligand.IL-21R-TCR-T showed better proliferation upon activation and superior antitumor function in vitro and in vivo.IL-21R-TCR-T exhibited a less differentiated,exhausted and apoptotic phenotype than conventional TCR-T upon repetitive tumor antigen stimulation.The novel IL-21 receptor in our study programs powerful TCR-T and can avoid side effects induced by IL-21 systemic utilization.The novel IL-21 receptor creates new opportunities for next-generation TCR-T against HCC.

Complementation of Wild-Type and Drug-Resistant Hepatitis B Virus Genomes to Maintain Viral Replication and Rescue Virion Production under Nucleos(t)ide Analogs

As the open reading frames of hepatitis B virus(HBV)genomes are overlapping,resistance mutations(MTs)in HBV polymerase may result in stop codon MTs in hepatitis B surface proteins,which are usually detected as a mixed population with wild-type(WT)HBV.The question was raised how the coexistence of nucleos(t)ide analogs(NAs)resistance MTs and WT sequences affects HBV replication.In the present study,HBV genomes with frequently detected reverse transcriptase(RT)/surface truncation MTs,rtA181T/sW172^*,rtV191I/sW182^*and rtM204I/sW196^*,were phenotypically characterized alone or together with their WT counterparts in different ratios by transient transfection in the absence or presence of Nas.In the absence of Nas,RT/surface truncation MTs impaired the expression and secretion of HBV surface proteins,and had a dose-dependent negative effect on WT HBV virion secretion.However,in the presence of Nas,coexistence of MTs with WT maintained viral replication,and the presence of WT was able to rescue the production of MT HBV virions.Our findings reveal that complementation of WT and MT HBV genomes is highly effective under drug treatment.

Targeting cIAPs,a New Option for Functional Cure of Chronic Hepatitis B Infection?

Hepatitis B virus (HBV)infection causes a wide spectrum of liver diseases in more than 240million people worldwide (Ott et al.2012).Patients with chronic hepatitis B (CHB)may progress to HBV-related end-stage liver diseases,such as hepatic failure,cirrhosis or hepatocellular carcinoma (HCC)(Tang et al.2018).

The IL-1R/TLR signaling pathway is essential for efficient CD8+ T-cell responses against hepatitis B virus in the hydrodynamic injection mouse model

The outcome of hepatitis B viral(HBV)infection is determined by the complex interactions between replicating HBV and the immune system.While the role of the adaptive immune system in the resolution of HBV infection has been studied extensively,the contribution of innate immune mechanisms remains to be defined.Here we examined the role of the interleukin-1 receptor/Toll-like receptor(IL-1R/TLR)signaling pathway in adaptive immune responses and viral clearance by exploring the HBV mouse model.Hydrodynamic injection with a replication-competent HBV genome was performed in wild-type mice(WT)and a panel of mouse strains lacking specific innate immunity component expression.We found higher levels of HBV protein production and replication in Tlr2^(−/−),Tlr23479^(−/−),3d/Tlr24^(−/−),Myd88/Trif^(−/−)and Irak4^(−/−)mice,which was associated with reduced HBV-specific CD8+T-cell responses in these mice.Importantly,HBV clearance was delayed for more than 2 weeks in 3d/Tlr24^(−/−),Myd88/Trif^(−/−)and Irak4^(−/−)mice compared to WT mice.HBV-specific CD8+T-cell responses were functionally impaired for producing the cytokines IFN-γ,TNF-αand IL-2 in TLR signaling-deficient mice compared to WT mice.In conclusion,the IL-1R/TLR signaling pathway might contribute to controlling HBV infection by augmenting HBV-specific CD8+T-cell responses.

Vertical characteristics and source identification of PM_(10) in Tianjin

Ambient PM 10 （particulate matter with a diameter less than 10μm） concentrations were measured on a 255 meter tower in Tianjin,China.The samples were collected at four vertical levels （10,40,120 and 220 m）.Vertical characteristics for PM 10 samples were studied.The results showed that the concentrations of PM 10 and constituent species had a negative correlation with the sampling height.The highest concentrations of PM 10 and species were obtained at the 10 m level,and the lowest concentrations were measured at the 220 m level.For the fractions of species to total mass,SO 4 2- and NO 3- had higher values （fraction） at greater height;while Ca had a higher fraction at lower height.Possible source categories for the PM 10 ambient dataset were identified by the principal component analysis method.The possible source categories included crustal dust,vehicles,cement dust,and incineration as well as secondary sulfate and nitrate sources.Analysis of meteorological factors on PM 10 concentrations indicated that wind speed and inversion may be the main factors contributing to different concentrations of PM 10 at different heights.

Viral and Antibody Kinetics of COVID-19 Patients with Different Disease Severities in Acute and Convalescent Phases:A 6-Month Follow-Up Study

Coronavirus disease 2019(COVID-19),caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),has spread rapidly around the world,posing a major threat to human health and the economy.Currently,long-term data on viral shedding and the serum antibody responses in COVID-19 patients are still limited.Herein,we report the clinical features,viral RNA loads,and serum antibody levels in a cohort of 112 COVID-19 patients admitted to the Honghu People’s Hospital,Hubei Province,China.Overall,5.36%(6/112)of patients showed persistent viral RNA shedding(>45 days).The peak viral load was higher in the severe disease group than in the mild group(median cycle threshold value,36.4 versus 31.5;P=0.002).For most patients the disappearance of IgM antibodies occurred approximately 4–6 weeks after symptoms onset,while IgG persisted for over 194 days after the onset of symptoms,although patients showed a 46%reduction in antibodies titres against SARS-CoV-2 nucleocapsid protein compared with the acute phase.We also studied18 asymptomatic individuals with RT-qPCR confirmed SARS-CoV-2 infection together with 17 symptomatic patients,and the asymptomatic individuals were the close contacts of these symptomatic cases.Delayed IgG seroconversion and lower IgM seropositive rates were observed in asymptomatic individuals.These data indicate that higher viral loads and stronger antibody responses are related to more severe disease status in patients with SARS-CoV-2 infection,and the antibodies persisted in the recovered patient for more than 6 months so that the vaccine may provide protection against SARS-CoV-2 infection.

Designing high-strength titanium alloy using pseudo-spinodal mechanism through diffusion multiple experiment and CALPHAD calculation

This study used the pseudo-spinodal mechanism to obtain the ultrafineαphase for designing highstrength titanium alloy.Diffusion multiple experiments were designed to find the composition range of TM-x Mo-y V alloy(TM:Ti-4.5 Al-2 Cr-2.5 Nb-2 Zr-1 Sn)for obtaining ultrafineαphase.CALPHAD results confirm that when the alloy composition is located near the intersection of theαandβphase free energy curves,the alloy will undergo pseudo-spinodal transformation and obtain the ultrafineαphase.The designed TM-6 Mo-3 V alloy has a yield strength of 1411 MPa and an elongation of 6.5%.The strength of the alloy depends on the thickness,fraction of theαphase and the solid solution strengthening effect of the alloying elements.The deformation mechanism of the alloy is the dislocation slip of theαandβphases and the twin deformation of theαphase.The large number ofα/βinterfaces produced by the fineαphase is the main reason for limiting the ductility of the alloy.The use of the pseudo-spinodal mechanism combined with diffusion multiple experiments and CALPHAD is an effective method for designing high-strength titanium alloys.

The dynamic responses of lamellar and equiaxed nearβ-Ti alloys subjected to multi-pass cross rolling

This work gives a comparison on the microstructural characteristics,textural discrepancies,and twinning behaviors of lamellar and equiaxed nearβ-Ti alloys during multi-pass cross rolling with a rolling reduction of 20%,50%and 80%.The results showed that the restoration mechanism of the alloy inβphase is strongly dependent on theαmorphologies,and in comparison,strain path has weaker influences on the grain refinement of theβmatrix.Therefore,the texture intensities of bothαandβphases were weakened owing to the dynamic recrystallization(DRX)of the two phases in the equiaxed microstructure.While,with regard to the lamellar microstructure,dynamic recovery(DRV)of theβphase predominated,forming elongatedβsubgrains.Besides,theαandβmatrix in lamellar microstructures obeyed the Burgers orientation relationship,which was gradually broken down until the final reduction.Lastly,the{1101}twinning exhibits a strong size effect.With the continuous DRX ofαphases,theα-twinning is suppressed owing to progressive grain refinement.The activation ofβ-twinning,namely{332}?113?and{112}?111?,in nearβ-Ti alloys is heavily dependent on the deficientβ-stabilizing elements and the local stress concentration.These findings provide an effective way to obtain ultra-fine grain microstructures of this alloy.

Analysis of crystallographic orientation and morphology of microstructure during hot working for an alpha/beta titanium alloy

This work focuses on analysis of microstructure morphology and crystallographic orientation for Ti-17 alloy during hot working.The results show that alpha phase and beta phase influence each other and there is a coordinate deformation between them.The non-uniform deformation is observed under small deformation conditions.The observing area can be divided into small deformation zone(area L)and large deformation zone(area H).Both alpha and beta phases remain the initial morphology,and they have better capability of coordinate deformation in area L,while coordinate capability is weak in area H in which alpha phase is globularized.Correspondingly,the Burgers orientation relations are well preserved in area L,but the orientation relations are more or less destroyed in area H.Dynamic recovery is the main mechanism of beta phase evolution when height reduction is lower.By contrast,the continuous dynamic recrystallization(CDRX)of beta phase gradually dominates the deformation pattern as the deformation increases.An uniformly globularized alpha structure is obtained under large deformation condition.The unsynchronized rotation of alpha phase around<11-20>pole occurs during deformation,which leads to the uniform crystal structure inside the same alpha lamellae.This process is an important step of globularization of the lamellar structure.

Noncanonical NF-κB Signaling Pathway in Liver Diseases

The noncanonical NF-κB signaling pathway is an important branch of NF-κB signaling.It is involved in regulating multiple important biological processes,including inflammation and host immune response.A central adaptor protein of the noncanonical NF-κB pathway is NF-κB-inducing kinase(NIK),which activates the downstream kinase IKKαto process p100 to p52,thereby forming the RelB/p52 heterodimer to initiate the expression of target genes.Currently,many specific inhibitors and monoclonal antibodies targeting or triggering this pathway are being developed and tested for various diseases,including cancers,autoimmune diseases,and virus infection.Given that aberrant activation of the noncanonical NF-κB pathway is frequently observed in various liver diseases,targeting this pathway may be a promising therapeutic strategy to alleviate liver inflammation.Moreover,activation of this pathway may contribute to the antiviral immune response and promote the clearance of persistent hepatotropic virus infection.Here,we review the role of the noncanonical NF-κB pathway in the occurrence and development of different liver diseases,and discuss the potency and application of modulating the noncanonical NF-κB pathway for treatment of these liver diseases.

One-step preparation of branched PEG functionalized AIE-active luminescent polymeric nanoprobes

The synthesis of amphiphilic aggregation-induced emission （ALE） dyes based organic nanoparticles has recently attracted in- creasing attention in the biomedical fields. These AlE dyes based nanoparticles could effectively overcome the aggregation caused quenching effect of conventional organic dyes, making them promising candidates for fabrication of ultrabright organic luminescent nanomaterials. In this work, AIE-active luminescent polymeric nanoparticles （4-NH2-PEG-TPE-E LPNs） were facilely fabricated through Michael addition reaction between tetraphenylethene acrylate （TPE-E） and 4-arm-poly（ethylene glycol）-amine （4-NH2-PEG） in rather mild ambient. The 4-NH2-PEG can not only endow these AlE-active LPNs good water dispersibility, but also provide functional groups for further conjugation reaction. The size, morphology and luminescent prop- erties of 4-NH2-PEG-TPE-E LPNs were characterized by a series of techniques in detail. Results suggested that these AlE-active LPNs showed spherical morphology with diameter about 100-200 nm. The obtained 4-NH2-PEG-TPE-E LPNs display high water dispersibility and strong fluorescence intensity because of their self assembly and AlE properties of TPE-E. Biological evaluation results demonstrated that 4-NH2-PEG-TPE-E LPNs showed negative toxicity toward cancer cells and good fluorescent imaging performance. All of these features make 4-NHz-PEG-TPE-E LPNs promising candidates for biolog- ical imaging and therapeutic applications.