探讨基于Caprini 风险评估模型进行 DVT风险评估，并根据风险程度分级实施预见性护理方案在中重度烧伤患者中的应用价值。方法 以2019年1至6月收治的40例中重度烧伤住院患者为对照组，实施烧伤患者常规护理及一般 DVT 预防措施；以 2019年7至12月收治的 42 例中重度烧伤住院患者为观察组，采用Caprini评估量表进行DVT风险分级，并实施相应的预见性护理。比较两组干预效果。结果观察组患者DVT疾病知识掌握程度、疾病健康行为均优于对照组，差异有统计学意义；观察组预见性护理措施落实率为92%高于对照组，差异有统计学意义；干预后观察组发生血液淤滞1例，对照组发生2例DVT及6例血流淤滞，观察组效果优于对照组，差异均有统计学意义。结论 基于Caprini量表DVT风险评估的预见性护理方案能够提高中重度烧伤住院患者DVT疾病知识掌握程度及预防措施落实率，降低DVT发生率，有效提高护理满意度。
探讨人皮下脂肪来源干细胞（human subcutaneous adipose-derived stem cells，hADSCs）局部移植对正畸源性牙根吸收（orthodontically induced root resorption，OIRR）的影响，为临床应用hADSCs抑制OIRR提供实验依据。
取40只8周龄雄性SD大鼠随机分为实验组和对照组，每组20只，建立大鼠右侧上颌第1磨牙近中牙正畸牙移动（orthodontic tooth movement，OTM）模型。实验组大鼠于建模第1、4、8、12天注射25 μL含2.5×105个hADSCs的细胞悬液，对照组注射25 μL PBS。在加力前及加力7、14 d后获取大鼠上颌模型，于加力7、14 d后两组各处死10只大鼠并取材。体式显微镜测量OTM距离，扫描电镜观察压力侧牙根形态及测定牙根吸收面积比，HE染色观察压力侧牙根吸收及牙周组织改建并计算牙根吸收指数，抗酒石酸酸性磷酸酶染色计数压力侧牙周组织破牙骨质细胞和破骨细胞数量。
两组OTM距离均随加力时间延长而增加（
hADSCs 局部移植可能通过降低大鼠OTM过程中破牙骨质细胞和破骨细胞的数量来减小牙根吸收面积和深度，从而抑制OIRR。
为了研究初始温度470K、初始压力0.1MPa下乙醇掺混对航空煤油层流火焰燃烧特性的影响，文中利用定容燃烧弹设备结合仿真进行对比分析。结果表明，乙醇的加入显著提升了航空煤油的层流火焰燃烧速度，但仿真结果与试验数据之间存在较大的偏差，在低当量比时尤为严重，需进一步优化机理。在此基础上通过敏感性分析，发现对层流火焰燃烧速度影响较大的4个基元反应，通过对4个基元反应的参数进行调整，修改得到新模型，该模型与试验值存在较好的吻合度。
沟金针虫是我国北方旱作农田的重要地下害虫。近年来华北地区大面积推行保护性耕作措施和作物秸秆粉碎还田，冬小麦+夏玉米一年两熟连续轮作种植，为沟金针虫创造了有利的取食和栖息的环境。地处华北北部的固城站2018—2019年秋、冬、春三季气温出现了冷暖交替，尤其最低气温显著偏高，诱发麦田沟金针虫爆发性发生为害。据春季麦田挖土调查，虫口密度最高达144头/m2；虫口重量最重达18.764g/m2。58个调查点达防治指标5头/m2占98.27%。拔节～收获期调查虫口密度：孕穗期最高、拔节期次之、收获期最低。老熟幼虫虫体最长34.68mm，最宽4.90mm，最长增长4.68mm，最宽增大0.90mm。冬小麦+夏玉米禾本科作物连作种植田间虫口密度达35.3～40.4头/m2，显著高于前茬大豆、玉米、冬小麦休闲地，且花生地、春玉米地比大豆地虫口密度高5倍多，虫口重量高10倍以上。成熟期虫害麦田测产：籽粒减产36.8%，虫口密度增加10头/m2籽粒减产率增加4.824%，虫口重量增加1g/m2籽粒减产率增加3.871%，植株虫害率增加10%籽粒减产率增加11.587%。
Plants recognize microbe-associated molecular patterns (MAMPs) to activate immune responses and defense priming to defend against pathogen infections. Transcriptional regulation of gene expression is crucial for plant immunity and is mediated by multiple factors, including DNA methylation. However, it remains unknown whether and how DNA demethylation contributes to immune responses in MAMP-triggered immunity. Here, we report that active DNA demethylation is required for MAMP-triggered immunity to bacterial pathogens. The rdd-2 triple mutant carrying mutations in ROS1, DML2, and DML3 that encode DNA glycosylases, which are key DNA demethylation enzymes, exhibits compromised immune responses triggered by the MAMPs flg22 and elf18. Genome-wide methylome analysis reveals that flg22 induces rapid and specific DNA demethylation in an RDD-dependent manner. The expression levels of salicylic acid signaling-related and phytoalexin biosynthesis-related genes are tightly associated with the flg22-induced promoter demethylation. The compromised accumulation of priming compounds and antimicrobial metabolites ultimately leads to a defense priming defect in the rdd-2 mutant. Our results reveal the critical role of active DNA demethylation in the MAMP-triggered immune response and provide unique insight into the molecular mechanism of flg22-modulated DNA demethylation.
Plants under pathogen attack produce high levels of the gaseous phytohormone ethylene to induce plant defense responses via the ethylene signaling pathway. The 1-aminocyclopropane-1-carboxylate synthase (ACS) is a critical rate-limiting enzyme of ethylene biosynthesis. Transcriptional and post-translational upregulation of ACS2 and ACS6 by the mitogen-activated protein kinases MPK3 and MPK6 are previously shown to be crucial for pathogen-induced ethylene biosynthesis in Arabidopsis. Here, we report that the fungal pathogen Botrytis cinerea-induced ethylene biosynthesis in Arabidopsis is under the negative feedback regulation by ethylene signaling pathway. The ethylene response factor ERF1A is further found to act downstream of ethylene signaling to negatively regulate the B. cinerea-induced ethylene biosynthesis via indirectly suppressing the expression of ACS2 and ACS6. Interestingly, ERF1A is shown to also upregulate defensin genes directly and therefore promote Arabidopsis resistance to B.cinerea. Furthermore, ERF1A is identified to be a substrate of MPK3 and MPK6, which phosphoactivate ERF1A to enhance its functions in suppressing ethylene biosynthesis and inducing defensin gene expression. Taken together, our data reveal that ERF1A and its phosphorylation by MPK3/MPK6 not only mediate the negative-feedback regulation of the B.cinerea-induced ethylene biosynthesis, but also upregulate defensin gene expression to increase Arabidopsis resistance to B. cinerea.
Pseudomonas syringae pv. actinidiae (Psa) causes bacterial canker, a devastating disease threatening the Actinidia fruit industry. In a search for non-host resistance genes against Psa, we find that the nucleotide-binding leucine-rich repeat receptor (NLR) protein ZAR1 from both Arabidopsis and Nicotiana benthamiana (Nb) recognizes HopZ5 and triggers cell death. The recognition requires ZED1 in Arabidopsis and JIM2 in Nb plants, which are members of the ZRK pseudokinases and known components of the ZAR1 resistosome. Surprisingly, Arabidopsis ZAR1 and RPM1, another NLR known to recognize HopZ5, confer disease resistance to HopZ5 in a strain-specific manner. Thus, ZAR1, but not RPM1, is solely required for resistance to P.s. maculicola ES4326 (Psm) carrying hopZ5, whereas RPM1 is primarily required for resistance to P.s. tomato DC3000 (Pst) carrying hopZ5. Furthermore, the ZAR1-mediated resistance to Psm hopZ5 in Arabidopsis is insensitive to SOBER1, which encodes a deacetylase known to suppress the RPM1-mediated resistance to Pst hopZ5. In addition, hopZ5 enhances P.syringae virulence in the absence of ZAR1 or RPM1 and that SOBER1 abolishes such virulence function. Together the study suggests that ZAR1 may be used for improving Psa resistance in Actinidia and uncovers previously unknown complexity of effector-triggered immunity and effector-triggered virulence.
Wild emmer wheat (Triticum dicoccoides, WEW) is an immediate progenitor of both the cultivated tetraploid and hexaploid wheats and it harbors rich genetic diversity against powdery mildew caused by Blumeria graminis f. sp. tritici (Bgt). A powdery mildew resistance gene MlIW172 originated from WEW accession IW172 (G-797-M) is fine mapped in a 0.048 centimorgan (cM) genetic interval on 7AL, corresponding to a genomic region spanning 233kb, 1Mb and 800kb in Chinese Spring, WEW Zavitan, and T.urartu G1812, respectively. MlIW172 encodes a typical NLR protein NLRIW172 and physically locates in an NBS-LRR gene cluster. NLR is subsequently identified as a new allele of Pm60, and its function is validated by EMS mutagenesis and transgenic complementation. Haplotype analysis of the Pm60 alleles reveals diversifications in sequence variation in the locus and presence and absence variations (PAV) in WEW populations. Four common single nucleotide variations (SNV) are detected between the Pm60 alleles from WEW and T.urartu, indicative of speciation divergence between the two different wheat progenitors. The newly identified Pm60 alleles and haplotypes in WEW are anticipated to be valuable for breeding powdery mildew resistance wheat cultivars via marker-assisted selection.
Plasma membrane H+-ATPases (PM H+-ATPases) are critical proton pumps that export protons from the cytoplasm to the apoplast. The resulting proton gradient and difference in electrical potential energize various secondary active transport events. PM H+-ATPases play essential roles in plant growth, development, and stress responses. In this review, we focus on recent studies of the mechanism of PM H+-ATPases in response to abiotic stresses in plants, such as salt and high pH, temperature, drought, light, macronutrient deficiency, acidic soil and aluminum stress, as well as heavy metal toxicity. Moreover, we discuss remaining outstanding questions about how PM H+-ATPases contribute to abiotic stress responses.
In response to dynamically altered environments, plants must finely coordinate the balance between growth and stress responses for their survival. However, the underpinning regulatory mechanisms remain largely elusive. The phytohormone gibberellin promotes growth via a derepression mechanism by proteasomal degradation of the DELLA transcription repressors. Conversely, the stress-induced burst of nitric oxide (NO) enhances stress tolerance, largely relying on NO-mediated S-nitrosylation, a redox-based posttranslational modification. Here, we show that S-nitrosylation of Cys-374 in the Arabidopsis RGA protein, a key member of DELLAs, inhibits its interaction with the F-box protein SLY1, thereby preventing its proteasomal degradation under salinity condition. The accumulation of RGA consequently retards growth but enhances salt tolerance. We propose that NO negatively regulates gibberellin signaling via S-nitrosylation of RGA to coordinate the balance of growth and stress responses when challenged by adverse environments.
Plants require solar energy to grow through oxygenic photosynthesis; however, when light intensity exceeds the optimal range for photosynthesis, it causes abiotic stress and physiological damage in plants. In response to high light stress, plants initiate a series of signal transduction from chloroplasts to whole cells and from locally stressed tissues to the rest of the plant body. These signals trigger a variety of physiological and biochemical reactions intended to mitigate the deleterious effects of high light intensity, such as photodamage and photoinhibition. Light stress protection mechanisms include chloroplastic Reactive oxygen species (ROS) scavenging, chloroplast and stomatal movement, and anthocyanin production. Photosynthetic apparatuses, being the direct targets of photodamage, have also developed various acclimation processes such as thermal energy dissipation through nonphotochemical quenching (NPQ), photorepair of Photosystem II (PSII), and transcriptional regulation of photosynthetic proteins. Fluctuating light is another mild but persistent type of light stress in nature, which unfortunately has been poorly investigated. Current studies, however, suggest that state transitions and cyclic electron transport are the main adaptive mechanisms for mediating fluctuating light stress in plants. Here, we review the current breadth of knowledge regarding physiological and biochemical responses to both high light stress and fluctuating light stress.
Salt stress adversely affects plant growth, development, and crop yield. Rice (Oryza sativa L.) is one of the most salt-sensitive cereal crops, especially at the early seedling stage. Mitogen-activated protein kinase (MAPK/MPK) cascades have been shown to play critical roles in salt response in Arabidopsis. However, the roles of the MPK cascade signaling in rice salt response and substrates of OsMPK remain largely unknown. Here, we report that the salt-induced OsMPK4-Ideal Plant Architecture 1 (IPA1) signaling pathway regulates the salt tolerance in rice. Under salt stress, OsMPK4 could interact with IPA1 and phosphorylate IPA1 at Thr180, leading to degradation of IPA1. Genetic evidence shows that IPA1 is a negative regulator of salt tolerance in rice, whereas OsMPK4 promotes salt response in an IPA1-dependent manner. Taken together, our results uncover an OsMPK4-IPA1 signal cascade that modulates the salt stress response in rice and sheds new light on the breeding of salt-tolerant rice varieties.
Eukaryotic cells are confined by membranes that create hydrophobic barriers for substance and information exchange between the inside and outside of the cell. These barriers are formed by assembly of lipids and protein in aqueous environments. Lipids not only serve as building blocks for membrane construction, but also possess regulatory functions in cellular activities. These regulatory lipids are non-uniformly distributed in membrane systems; their temporal and spatial accumulation in specific membranes decodes environmental cues and changes cellular activity accordingly. Phosphoinositides (PIs) are phospholipids that exert regulatory effects. In recent years, research on PIs roles in regulating plant growth, development, and responses to environmental stress is increasing. Several reviews have been published on the composition of PIs, intermolecular transferring of PIs by lipid kinases (phosphatases) or PI-PLCs, subcellular localization, and specially their functions in plant developments. Herein, we review the crucial regulatory functions of PIs in plant stress responses, with a particular focus on PIs involved in membrane trafficking.
Phenolamide (PA) metabolites play important roles in the interaction between plants and pathogens. The putrescine hydroxycinnamoyl transferase genes OsPHT3 and OsPHT4 positively regulate rice cell death and resistance to Magnaporthe oryzae. The bZIP transcription factor APIP5, a negative regulator of cell death and rice immunity, directly binds to the OsPHT4 promoter to regulate putrescine-derived PAs. Whether other hydroxycinnamoyl transferase (HT) genes also participate in APIP5-mediated immunity remains unclear. Surprisingly, we find that genes encoding agmatine hydroxycinnamoyl transferases OsAHT1 and OsAHT2, tryptamine hydroxycinnamoyl transferases OsTBT1 and OsTBT2, and tyramine hydroxycinnamoyl transferases OsTHT1 and OsTHT2, responsible for the biosynthesis of polyamine-derived PAs are all up-regulated in APIP5-RNAi transgenic plants compared with segregated wild-type rice. Furthermore, both OsAHT1/2 and OsTBT1/2 are induced during M.oryzae infection, showing expression patterns similar to those previously reported for OsTHT1/2 and OsPHT3/4. Transgenic plants overexpressing either OsAHT2-GFP or OsTBT1-GFP show enhanced resistance against M.oryzae and accumulated more PA metabolites and lignin compared with wild-type plants. Interestingly, as demonstrated for OsPHT4, APIP5 directly binds to the promoters of OsAHT1/2, OsTBT1/2, and OsTHT1/2, repressing their transcription. Together, these results indicate that the HT genes are common targets of APIP5 and that PAs play critical roles in rice immunity.
Hydrogen sulfide (H2S) was once principally considered the perpetrator of plant growth cessation and cell death. However, this has become an antiquated view, with cumulative evidence showing that the H2S serves as a biological signaling molecule notably involved in abiotic stress response and adaptation, such as defense by phytohormone activation, stomatal movement, gene reprogramming, and plant growth modulation. Reactive oxygen species (ROS)-dependent oxidative stress is involved in these responses. Remarkably, an ever-growing body of evidence indicates that H2S can directly interact with ROS processing systems in a redox-dependent manner, while it has been gradually recognized that H2S-based posttranslational modifications of key protein cysteine residues determine stress responses. Furthermore, the reciprocal interplay between H2S and nitric oxide (NO) in regulating oxidative stress has significant importance. The interaction of H2S with NO and ROS during acclimation to abiotic stress may vary from synergism to antagonism. However, the molecular pathways and factors involved remain to be identified. This review not only aims to provide updated information on H2S action in regulating ROS-dependent redox homeostasis and signaling, but also discusses the mechanisms of H2S-dependent regulation in the context of oxidative stress elicited by environmental cues.
Plant diseases caused by diverse pathogens lead to a serious reduction in crop yield and threaten food security worldwide. Genetic improvement of plant immunity is considered as the most effective and sustainable approach to control crop diseases. In the last decade, our understanding of plant immunity at both molecular and genomic levels has improved greatly. Combined with advances in biotechnologies, particularly clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9-based genome editing, we can now rapidly identify new resistance genes and engineer disease-resistance crop plants like never before. In this review, we summarize the current knowledge of plant immunity and outline existing and new strategies for disease resistance improvement in crop plants. We also discuss existing challenges in this field and suggest directions for future studies.
The phytohormone jasmonate plays a pivotal role in various aspects of plant life, including developmental programs and defense against pests and pathogens. A large body of knowledge on jasmonate biosynthesis, signal transduction as well as its functions in diverse plant processes has been gained in the past two decades. In addition, there exists extensive crosstalk between jasmonate pathway and other phytohormone pathways, such as salicylic acid (SA) and gibberellin (GA), in co-regulation of plant immune status, fine-tuning the balance of plant growth and defense, and so on, which were mostly learned from studies in the dicotyledonous model plants Arabidopsis thaliana and tomato but much less in monocot. Interestingly, existing evidence suggests both conservation and functional divergence in terms of core components of jasmonate pathway, its biological functions and signal integration with other phytohormones, between monocot and dicot. In this review, we summarize the current understanding on JA signal initiation, perception and regulation, and highlight the distinctive characteristics in different lineages of plants.
The obstruction of post-insulin receptor signaling is the main mechanism of insulin-resistant diabetes. Progestin and adipoQ receptor 3 (PAQR3), a key regulator of inflammation and metabolism, can negatively regulate the PI3K/AKT signaling pathway. Here, we report that gentiopicroside (GPS), the main bioactive secoiridoid glycoside of Gentiana manshurica Kitagawa, decreased lipid synthesis and increased glucose utilization in palmitic acid (PA) treated HepG2 cells. Additionally, GPS improved glycolipid metabolism in streptozotocin (STZ) treated high-fat diet (HFD)-induced diabetic mice. Our findings revealed that GPS promoted the activation of the PI3K/AKT axis by facilitating DNA-binding protein 2 (DDB2)-mediated PAQR3 ubiquitinated degradation. Moreover, results of surface plasmon resonance (SPR), microscale thermophoresis (MST) and thermal shift assay (TSA) indicated that GPS directly binds to PAQR3. Results of molecular docking and cellular thermal shift assay (CETSA) revealed that GPS directly bound to the amino acids of the PAQR3 NH2-terminus including Leu40, Asp42, Glu69, Tyr125 and Ser129, and spatially inhibited the interaction between PAQR3 and the PI3K catalytic subunit (P110α) to restore the PI3K/AKT signaling pathway. In summary, our study identified GPS, which inhibits PAQR3 expression and directly targets PAQR3 to restore insulin signaling pathway, as a potential drug candidate for the treatment of diabetes.
Lipid nanoparticle (LNP) is commonly used to deliver mRNA vaccines. Currently, LNP optimization primarily relies on screening ionizable lipids by traditional experiments which consumes intensive cost and time. Current study attempts to apply computational methods to accelerate the LNP development for mRNA vaccines. Firstly, 325 data samples of mRNA vaccine LNP formulations with IgG titer were collected. The machine learning algorithm, lightGBM, was used to build a prediction model with good performance (R2 > 0.87). More importantly, the critical substructures of ionizable lipids in LNPs were identified by the algorithm, which well agreed with published results. The animal experimental results showed that LNP using DLin-MC3-DMA (MC3) as ionizable lipid with an N/P ratio at 6:1 induced higher efficiency in mice than LNP with SM-102, which was consistent with the model prediction. Molecular dynamic modeling further investigated the molecular mechanism of LNPs used in the experiment. The result showed that the lipid molecules aggregated to form LNPs, and mRNA molecules twined around the LNPs. In summary, the machine learning predictive model for LNP-based mRNA vaccines was first developed, validated by experiments, and further integrated with molecular modeling. The prediction model can be used for virtual screening of LNP formulations in the future.
Photothermal therapy has been intensively investigated for treating cancer in recent years. However, the long-term therapeutic outcome remains unsatisfying due to the frequently occurred metastasis and recurrence. To address this challenge, immunotherapy has been combined with photothermal therapy to activate anti-tumor immunity and relieve the immunosuppressive microenvironment within tumor sites. Here, we engineered silica-based core-shell nanoparticles (JQ-1@PSNs-R), in which silica cores were coated with the photothermal agent polydopamine, and a bromodomain-containing protein 4 (BRD4) inhibitor JQ-1 was loaded in the polydopamine layer to combine photothermal and immune therapy for tumor elimination. Importantly, to improve the therapeutic effect, we increased the surface roughness of the nanoparticles by hydrofluoric acid (HF) etching during the fabrication process, and found that the internalization of JQ-1@PSNs-R was significantly improved, leading to a strengthened photothermal killing effect as well as the increased intracellular delivery of JQ-1. In the animal studies, the multifunctional nanoparticles with rough surfaces effectively eradicated melanoma via photothermal therapy, successfully activated tumor-specific immune responses against residual tumor cells, and further prevented tumor metastasis and recurrence. Our results indicated that JQ-1@PSNs-R could serve as an innovative and effective strategy for combined cancer therapy.
The p21 activated kinase 4 (PAK4) is serine/threonine protein kinase that is critical for cancer progression. Guided by X-ray crystallography and structure-based optimization, we report a novel subseries of C-3-substituted 6-ethynyl-1H-indole derivatives that display high potential and specificity towards group II PAKs. Among these inhibitors, compound 55 exhibited excellent inhibitory activity and kinase selectivity, displayed superior anti-migratory and anti-invasive properties against the lung cancer cell line A549 and the melanoma cell line B16. Compound 55 exhibited potent in vivo antitumor metastatic efficacy, with over 80% and 90% inhibition of lung metastasis in A549 or B16-BL6 lung metastasis models, respectively. Further mechanistic studies demonstrated that compound 55 mitigated TGF-β1-induced epithelial-mesenchymal transition (EMT).
Tripterygium wilfordii is a valuable medicinal plant rich in biologically active diterpenoids, but there are few studies on the origins of these diterpenoids in its secondary metabolism. Here, we identified three regions containing tandemly duplicated diterpene synthase genes on chromosomes (Chr) 17 and 21 of T. wilfordii and obtained 11 diterpene synthases with different functions. We further revealed that these diterpene synthases underwent duplication and rearrangement at approximately 2.3–23.7 million years ago (MYA) by whole-genome triplication (WGT), transposon mediation, and tandem duplication, followed by functional divergence. We first demonstrated that four key amino acids in the sequences of TwCPS3, TwCPS5, and TwCPS6 were altered during evolution, leading to their functional divergence and the formation of diterpene secondary metabolites. Then, we demonstrated that the functional divergence of three TwKSLs was driven by mutations in two key amino acids. Finally, we discovered the mechanisms of evolution and pseudogenization of miltiradiene synthases in T. wilfordii and elucidated that the new function in TwMS1/2 from the terpene synthase (TPS)-b subfamily was caused by progressive changes in multiple amino acids after the WGT event. Our results provide key evidence for the formation of diverse diterpenoids during the evolution of secondary metabolites in T. wilfordii.
PD-1 and PD-L1 antibodies have brought about extraordinary clinical benefits for cancer patients, and their indications are expanding incessantly. Currently, most PD-1/PD-L1 agents are administered intravenously, which may be uncomfortable for some cancer patients. Herein, we develop a novel oral-delivered small molecular, YPD-29B, which specifically targets human PD-L1. Our data suggested that YPD-29B could potently and selectively block the interaction between PD-L1 and PD-1, but did not inhibit any other immune checkpoints. Mechanistically, YPD-29B induced human PD-L1 dimerization and internalization, which subsequently activated T lymphocytes and therefore overcomes immunity tolerance in vitro. YDP-29B was modified as the YPD-30 prodrug to improve druggability. Using humanized mice with human PD-1 xenografts of human PD-L1 knock-in mouse MC38 cancer cells, we demonstrated that YPD-30 exhibited significant antitumor activity and was well tolerated in vivo. Taken together, our results indicate that YPD-30 serves as a promising therapeutic candidate for anti-human PD-L1 cancer immunotherapy.
Cyclooxygenases play a vital role in inflammation and are responsible for the production of prostaglandins. Two cyclooxygenases are described, the constitutive cyclooxygenase-1 and the inducible cyclooxygenase-2, for which the target inhibitors are the non-steroidal anti-inflammatory drugs (NSAIDs). Prostaglandins are a class of lipid compounds that mediate acute and chronic inflammation. NSAIDs are the most frequent choices for treatment of inflammation. Nevertheless, currently used anti-inflammatory drugs have become associated with a variety of adverse effects which lead to diminished output even market withdrawal. Recently, more studies have been carried out on searching novel selective COX-2 inhibitors with safety profiles. In this review, we highlight the various structural classes of organic and natural scaffolds with efficient COX-2 inhibitory activity reported during 2011–2021. It will be valuable for pharmaceutical scientists to read up on the current chemicals to pave the way for subsequent research.
Hematopoietic stem cell (HSC) transplantation is the only curative therapy for many diseases. HSCs from umbilical cord blood (UCB) source have many advantages over from bone marrow. However, limited HSC dose in a single CB unit restrict its widespread use. Over the past two decades, ex vivo HSC expansion with small molecules has been an effective approach for obtaining adequate HSCs. Till now, several small-molecule compounds have entered the phase I/II trials, showing safe and favorable pharmacological profiles. As HSC expansion has become a hot topic over recent years, many newly identified small molecules along with novel biological mechanisms for HSC expansion would help solve this challenging issue. Here, we will give an overview of HSC biology, discovery and medicinal chemistry development of small molecules, natural products targeting for HSC expansion, and their recent clinical progresses, as well as potential protein targets for HSC expansion.
Nonalcoholic fatty liver disease (NAFLD) is characterized by hepatic steatosis and insulin resistance and there are currently no approved drugs for its treatment. Hyperactivation of mTOR complex 1 (mTORC1) and subsequent impairment of the transcription factor EB (TFEB)-mediated autophagy–lysosomal pathway (ALP) are implicated in the development of NAFLD. Accordingly, agents that augment hepatic TFEB transcriptional activity may have therapeutic potential against NAFLD. The objective of this study was to investigate the effects of nuciferine, a major active component from lotus leaf, on NAFLD and its underlying mechanism of action. Here we show that nuciferine activated ALP and alleviated steatosis, insulin resistance in the livers of NAFLD mice and palmitic acid-challenged hepatocytes in a TFEB-dependent manner. Mechanistic investigation revealed that nuciferine interacts with the Ragulator subunit hepatitis B X-interacting protein and impairs the interaction of the Ragulator complex with Rag GTPases, thereby suppressing lysosomal localization and activity of mTORC1, which activates TFEB-mediated ALP and further ameliorates hepatic steatosis and insulin resistance. Our present results indicate that nuciferine may be a potential agent for treating NAFLD and that regulation of the mTORC1–TFEB–ALP axis could represent a novel pharmacological strategy to combat NAFLD.
Algae are a large group of photosynthetic organisms responsible for approximately half of the earth's total photosynthesis. In addition to their fundamental ecological roles as oxygen producers and as the food base for almost all aquatic life, algae are also a rich source of bioactive natural products, including several clinical drugs. Cytochrome P450 enzymes (P450s) are a superfamily of biocatalysts that are extensively involved in natural product biosynthesis by mediating various types of reactions. In the post-genome era, a growing number of P450 genes have been discovered from algae, indicating their important roles in algal life-cycle. However, the functional studies of algal P450s remain limited. Benefitting from the recent technical advances in algae cultivation and genetic manipulation, the researches on P450s in algal natural product biosynthesis have been approaching to a new stage. Moreover, some photoautotrophic algae have been developed into “photo-bioreactors” for heterologous P450s to produce high-value added pharmaceuticals and chemicals in a carbon-neutral or carbon-negative manner. Here, we comprehensively review these advances of P450 studies in algae from 2000 to 2021.
Nicotinamide phosphoribosyl transferase (NAMPT) is considered as a promising target for cancer therapy given its critical engagement in cancer metabolism and inflammation. However, therapeutic benefit of NAMPT enzymatic inhibitors appears very limited, likely due to the failure to intervene non-enzymatic functions of NAMPT. Herein, we show that NAMPT dampens antitumor immunity by promoting the expansion of tumor infiltrating myeloid derived suppressive cells (MDSCs) via a mechanism independent of its enzymatic activity. Using proteolysis-targeting chimera (PROTAC) technology, PROTAC A7 is identified as a potent and selective degrader of NAMPT, which degrades intracellular NAMPT (iNAMPT) via the ubiquitin–proteasome system, and in turn decreases the secretion of extracellular NAMPT (eNAMPT), the major player of the non-enzymatic activity of NAMPT. In vivo, PROTAC A7 efficiently degrades NAMPT, inhibits tumor infiltrating MDSCs, and boosts antitumor efficacy. Of note, the anticancer activity of PROTAC A7 is superior to NAMPT enzymatic inhibitors that fail to achieve the same impact on MDSCs. Together, our findings uncover the new role of enzymatically-independent function of NAMPT in remodeling the immunosuppressive tumor microenvironment, and reports the first NAMPT PROTAC A7 that is able to block the pro-tumor function of both iNAMPT and eNAMPT, pointing out a new direction for the development of NAMPT-targeted therapies.
Cancer immunotherapy is impaired by the intrinsic and adaptive immune resistance. Herein, a bispecific prodrug nanoparticle was engineered for circumventing immune evasion of the tumor cells by targeting multiple immune resistance mechanisms. A disulfide bond-linked bispecific prodrug of NLG919 and JQ1 (namely NJ) was synthesized and self-assembled into a prodrug nanoparticle, which was subsequently coated with a photosensitizer-modified and tumor acidity-activatable diblock copolymer PHP for tumor-specific delivery of NJ. Upon tumor accumulation via passive tumor targeting, the polymeric shell was detached for facilitating intracellular uptake of the bispecific prodrug. NJ was then activated inside the tumor cells for releasing JQ1 and NLG919 via glutathione-mediated cleavage of the disulfide bond. JQ1 is a bromodomain-containing protein 4 inhibitor for abolishing interferon gamma-triggered expression of programmed death ligand 1. In contrast, NLG919 suppresses indoleamine-2,3-dioxygenase 1-mediated tryptophan consumption in the tumor microenvironment, which thus restores robust antitumor immune responses. Photodynamic therapy (PDT) was performed to elicit antitumor immunogenicity by triggering immunogenic cell death of the tumor cells. The combination of PDT and the bispecific prodrug nanoparticle might represent a novel strategy for blockading multiple immune evasion pathways and improving cancer immunotherapy.
Neurodegenerative diseases (NDDs) such as Alzheimer's disease (AD) and Parkinson's disease (PD) are a heterogeneous group of disorders characterized by progressive degeneration of neurons. NDDs threaten the lives of millions of people worldwide and regretfully remain incurable. It is well accepted that dysfunction of mitochondria underlies the pathogenesis of NDDs. Dysfunction of mitochondria results in energy depletion, oxidative stress, calcium overloading, caspases activation, which dominates the neuronal death of NDDs. Therefore, mitochondria are the preferred target for intervention of NDDs. So far various mitochondria-targeting drugs have been developed and delightfully some of them demonstrate promising outcome, though there are still some obstacles such as targeting specificity, delivery capacity hindering the drugs development. In present review, we will elaborately address 1) the strategy to design mitochondria targeting drugs, 2) the rescue mechanism of respective mitochondria targeting drugs, 3) how to evaluate the therapeutic effect. Hopefully this review will provide comprehensive knowledge for understanding how to develop more effective drugs for the treatment of NDDs.
Remodeling the tumor microenvironment through reprogramming tumor-associated macrophages (TAMs) and increasing the immunogenicity of tumors via immunogenic cell death (ICD) have been emerging as promising anticancer immunotherapy strategies. However, the heterogeneous distribution of TAMs in tumor tissues and the heterogeneity of the tumor cells make the immune activation challenging. To overcome these dilemmas, a hybrid bacterium with tumor targeting and penetration, TAM polarization, and photothermal conversion capabilities is developed for improving antitumor immunotherapy in vivo. The hybrid bacteria (B.b@QDs) are prepared by loading Ag2S quantum dots (QDs) on the Bifidobacterium bifidum (B.b) through electrostatic interactions. The hybrid bacteria with hypoxia targeting ability can effectively accumulate and penetrate the tumor tissues, enabling the B.b to fully contact with the TAMs and mediate their polarization toward M1 phenotype to reverse the immunosuppressive tumor microenvironment. It also enables to overcome the intratumoral heterogeneity and obtain abundant tumor-associated antigens by coupling tumor penetration of the B.b with photothermal effect of the QDs, resulting in an enhanced immune effect. This strategy that combines B.b-triggered TAM polarization and QD-induced ICD achieved a remarkable inhibition of tumor growth in orthotopic breast cancer.
Breast cancer has become the most commonly diagnosed cancer type in the world. A combination of chemotherapy and photothermal therapy (PTT) has emerged as a promising strategy for breast cancer therapy. However, the intricacy of precise delivery and the ability to initiate drug release in specific tumor sites remains a challenging puzzle. Therefore, to ensure that the therapeutic agents are synchronously delivered to the tumor site for their synergistic effect, a multifunctional nanoparticle system (PCRHNs) is developed, which is grafted onto the prussian blue nanoparticles (PB NPs) by reduction-responsive camptothecin (CPT) prodrug copolymer, and then modified with tumor-targeting peptide cyclo(Asp-d-Phe-Lys-Arg-Gly) (cRGD) and hyaluronic acid (HA). PCRHNs exhibited nano-sized structure with good monodispersity, high load efficiency of CPT, triggered CPT release in response to reduction environment, and excellent photothermal conversion under laser irradiation. Furthermore, PCRHNs can act as a photoacoustic imaging contrast agent-guided PTT. In vivo studies indicate that PCRHNs exhibited excellent biocompatibility, prolonged blood circulation, enhanced tumor accumulation, allow tumor-specific chemo-photothermal therapy to achieve synergistic antitumor effects with reduced systemic toxicity. Moreover, hyperthermia-induced upregulation of heat shock protein 70 in the tumor cells could be inhibited by CPT. Collectively, PCRHNs may be a promising therapeutic way for breast cancer therapy.
Immunogenic cell death (ICD) plays a major role in cancer immunotherapy by stimulating specific T cell responses and restoring the antitumor immune system. However, effective type II ICD inducers without biotoxicity are still very limited. Herein, a tentative drug- or photosensitizer-free strategy was developed by employing enzymatic self-assembly of the peptide F-pY-T to induce mitochondrial oxidative stress in cancer cells. Upon dephosphorylation catalyzed by alkaline phosphatase overexpressed on cancer cells, the peptide F-pY-T self-assembled to form nanoparticles, which were subsequently internalized. These affected the morphology of mitochondria and induced serious reactive oxygen species production, causing the ICD characterized by the release of danger-associated molecular patterns (DAMPs). DAMPs enhanced specific immune responses by promoting the maturation of DCs and the intratumoral infiltration of tumor-specific T cells to eradicate tumor cells. The dramatic immunotherapeutic capacity could be enhanced further by combination therapy of F-pY-T and anti-PD-L1 agents without visible biotoxicity in the main organs. Thus, our results revealed an alternative strategy to induce efficient ICD by physically promoting mitochondrial oxidative stress.
Drug metabolism and pharmacokinetics (DMPK) is an important branch of pharmaceutical sciences. The nature of ADME (absorption, distribution, metabolism, excretion) and PK (pharmacokinetics) inquiries during drug discovery and development has evolved in recent years from being largely descriptive to seeking a more quantitative and mechanistic understanding of the fate of drug candidates in biological systems. Tremendous progress has been made in the past decade, not only in the characterization of physiochemical properties of drugs that influence their ADME, target organ exposure, and toxicity, but also in the identification of design principles that can minimize drug-drug interaction (DDI) potentials and reduce the attritions. The importance of membrane transporters in drug disposition, efficacy, and safety, as well as the interplay with metabolic processes, has been increasingly recognized. Dramatic increases in investments on new modalities beyond traditional small and large molecule drugs, such as peptides, oligonucleotides, and antibody-drug conjugates, necessitated further innovations in bioanalytical and experimental tools for the characterization of their ADME properties. In this review, we highlight some of the most notable advances in the last decade, and provide future perspectives on potential major breakthroughs and innovations in the translation of DMPK science in various stages of drug discovery and development.
Hybrid lipid-nanoparticle complexes have shown attractive characteristics as drug carriers due to their integrated advantages from liposomes and nanoparticles. Here we developed a kind of lipid-small molecule hybrid nanoparticles (LPHNPs) for imaging and treatment in an orthotopic glioma model. LPHNPs were prepared by engineering the co-assembly of lipids and an amphiphilic pheophorbide a-quinolinium conjugate (PQC), a mitochondria-targeting small molecule. Compared with the pure nanofiber self-assembled by PQC, LPHNPs not only preserve the comparable antiproliferative potency, but also possess a spherical nanostructure that allows the PQC molecules to be administrated through intravenous injection. Also, this co-assembly remarkably improved the drug-loading capacity and formulation stability against the physical encapsulation using conventional liposomes. By integrating the advantages from liposome and PQC molecule, LPHNPs have minimal system toxicity, enhanced potency of photodynamic therapy (PDT) and visualization capacities of drug biodistribution and tumor imaging. The hybrid nanoparticle demonstrates excellent curative effects to significantly prolong the survival of mice with the orthotopic glioma. The unique co-assembly of lipid and small molecule provides new potential for constructing new liposome-derived nanoformulations and improving cancer treatment.
Photothermal therapy has the characteristics of minimal invasiveness, controllability, high efficiency, and strong specificity, which can effectively make up for the toxic side effects and tumor resistance caused by traditional drug treatment. However, due to the limited tissue penetration of infrared light, it is difficult to promote and apply in clinical practice. The eye is the only transparent tissue in human, and infrared light can easily penetrate the eye tissue, so it is expected that photothermal therapy can be used to treat fundus diseases. Here in, a new nano-platform assembled by liposome and indocyanine green (ICG) was used to treat retinoblastoma. ICG was assembled in liposomes to overcome some problems of ICG itself. For example, ICG is easily quenched, self-aggregating and instability. Moreover, liposomes can prevent free ICG from being cleared through the systemic circulation. The construction of the nano-platform not only ensured the stability of ICG in vivo, but also realized imaging-guide photothermal therapy, which created a new strategy for the treatment of retinoblastoma.
Accurately delineating tumor boundaries is key to predicting survival rates of cancer patients and assessing response of tumor microenvironment to various therapeutic techniques such as chemotherapy and radiotherapy. This review discusses various strategies that have been deployed to accurately delineate tumor boundaries with particular emphasis on the potential of chemotherapeutic nanomaterials in tumor boundary delineation. It also compiles the types of tumors that have been successfully delineated by currently available strategies. Finally, the challenges that still abound in accurate tumor boundary delineation are presented alongside possible perspective strategies to either ameliorate or solve the problems. It is expected that the information communicated herein will form the first compendious baseline information on tumor boundary delineation with chemotherapeutic nanomaterials and provide useful insights into future possible paths to advancing current available tumor boundary delineation approaches to achieve efficacious tumor therapy.
Lipid nanoparticle (LNP)-based drug delivery systems have become the most clinically advanced non-viral delivery technology. LNPs can encapsulate and deliver a wide variety of bioactive agents, including the small molecule drugs, proteins and peptides, and nucleic acids. However, as the physicochemical properties of small- and macromolecular cargos can vary drastically, every LNP carrier system needs to be carefully tailored in order to deliver the cargo molecules in a safe and efficient manner. Our group applied the combinatorial library synthesis approach and in vitro and in vivo screening strategy for the development of LNP delivery systems for drug delivery. In this Review, we highlight our recent progress in the design, synthesis, characterization, evaluation, and optimization of combinatorial LNPs with novel structures and properties for the delivery of small- and macromolecular therapeutics both in vitro and in vivo. These delivery systems have enormous potentials for cancer therapy, antimicrobial applications, gene silencing, genome editing, and more. We also discuss the key challenges to the mechanistic study and clinical translation of new LNP-enabled therapeutics.
Glioma is a primary aggressive brain tumor with high recurrence rate. The poor efficiency of chemotherapeutic drugs crossing the blood-brain barrier (BBB) is well-known as one of the main challenges for anti-glioma therapy. Moreover, massive infiltrated tumor-associated macrophages (TAMs) in glioma further thwart the drug efficacy. Herein, a therapeutic nanosystem (SPP-ARV-825) is constructed by incorporating the BRD4-degrading proteolytic targeting chimera (PROTAC) ARV-825 into the complex micelle (SPP) composed of substance P (SP) peptide-modified poly(ethylene glycol)-poly(d,l-lactic acid)(SP-PEG-PDLLA) and methoxy poly(ethylene glycol)-poly(d,l-lactic acid) (mPEG-PDLLA, PP), which could penetrate BBB and target brain tumor. Subsequently, released drug engenders antitumor effect via attenuating cells proliferation, inducing cells apoptosis and suppressing M2 macrophages polarization through the inhibition of IRF4 promoter transcription and phosphorylation of STAT6, STAT3 and AKT. Taken together, our work demonstrates the versatile role and therapeutic efficacy of SPP-ARV-825 micelle against glioma, which may provide a novel strategy for glioma therapy in future.
沉积岩中的微量元素对沉积环境变化有较高的敏感度,是研究古沉积环境的有效手段。滇中盆地倒石头组是一套富锂的黏土岩地层,其古环境的研究对恢复该时期盆地沉积格局和锂元素富集具有重要意义。基于滇中盆地倒石头组两个典型钻孔样品详细的地球化学研究,探讨了倒石头组富锂黏土岩形成时的沉积环境及其对锂元素富集的影响。研究结果表明：所有样品Sr、Ga元素含量及Sr/Ba值指示研究区古水体介质为淡水陆相沉积环境；其
本文旨在筛选一株可拮抗中国大鲵源嗜水气单胞菌的后生元菌株，并对其进行抑菌特性的研究和细菌素基因簇的挖掘。以中国大鲵病原性嗜水气单胞菌（
随着现代法治体系的建立和不断完善，以及社会各项制度的成熟，以政教分离作为基本立原则的美国，在处理宗教信仰自由权问题上不断彰显了司法的能动性，最典型案件就是威斯康星州诉约德案。这场案件表面上是阿米什人的宗教信仰自由的权利和政府实施强制义务教育的权力之争、传统宗教教育和现代国民义务教育之争，实际上这场案件背后代表了民族传统文化与现代主流文明之间产生了巨大的张力、宗教信仰的神圣性与现代文明的世俗性之间也产生了巨大张力。本文将以威斯康星州诉约德案为例，从现代法律与宗教信仰自由之间的冲突为视角，探讨现代法治体系下的神圣与世俗之间的张力，以及在此基础上延伸出来的民族传统文化与现代主流文明之间的张力。
【目的】深入分析本土 4A 广告公司的计算化转型策略，以期进一步掌握广告业发展动态，为本土 4A 广告公司转型之路提供借鉴。【方法】通过案例分析法，对蓝色光标、省广集团等本土 4A 广告公司的经营范围、管理结构、技术应用、媒介投放、数据来源等进行研究总结。【结果】各本土 4A 广告公司应通过信息来源多元化、经营范围多样化、管理结构扁平化、内容形式细分化、媒介选择移动化等多方面进行计算化转型策略的尝试。【结论】向计算化转型将成为本土 4A 广告公司未来长期的发展趋势，也逐渐走向全面化、深层次、融合化。在此过程中如何处理创意与技术的关系将仍是本土 4A 广告公司需要解决的主要问题。
分析2018 — 2020年上海市分离的B/Victoria系流感毒株与疫苗株的匹配性及基因变异情况。
利用血凝抑制实验，对142株B/Victoria系流感毒株进行了抗原性分析，并选取了63株开展了血凝素（HA）及神经氨酸酶（NA）基因序列分析。
2018 — 2020年上海市流行的B/Victoria系流感毒株主要属于V1A.3分支，少数属于V1A.1分支及未缺失的V1A分支；抗原性分析显示，26.76%的流行株是疫苗株B/Colorado/06/2017鸡胚株的低反应株，与疫苗株相比，V1A.3分支流行株在HA蛋白的120环、150环、160环及190螺旋等关键的抗原决定簇及受体结合关键部位发生了5个氨基酸位点改变。
2018 — 2020监测年度B/Victoria系流行株与世界卫生组织推荐的疫苗株组分B/Colorado/06/2017匹配性不佳，仍需密切监测流行株的谱系及变异情况，为疫苗株的筛选提供可靠的数据。
俯卧位通气（PPV）作为机械通气治疗的一部分越来越受重视，相关共识和指南相继出台。俯卧位治疗（PPT）在烧伤临床诊疗中应用较为普遍。与传统意义的PPV相比，烧伤PPT在适应证、流程细节、注意事项等方面存在显著差异。因此，中国老年医学学会烧创伤分会和中华医学会烧伤外科学分会重症学组汇总循证证据，牵头制订了《成人烧伤俯卧位治疗全国专家共识（2022版）》，从PPT作用机制、适应证、使用流程等方面形成推荐意见，以供临床参考。
制备含氧化石墨烯（GO）的甲基丙烯酸酐化明胶（GelMA）水凝胶并探讨原位光聚合GO-GelMA复合水凝胶对小鼠全层皮肤缺损创面血管化的影响。
采用实验研究方法。将0.2 mg/mL的GO溶液50 μL均匀涂抹于导电胶上，烘干后于场发射扫描电子显微镜下观察GO的结构和大小。将人皮肤成纤维细胞（HSF）分为采用相应终质量浓度GO处理的0 μg/mL GO（不加GO溶液，下同）组、0.1 μg/mL GO组、1.0 μg/mL GO组、5.0 μg/mL GO组、10.0 μg/mL GO组，用酶标仪检测细胞培养48 h的吸光度值，以此表示细胞增殖活性（样本数为6）。将HSF和人脐静脉血管内皮细胞（HUVEC）分别分为采用相应终质量浓度GO处理的0 μg/mL GO组、0.1 μg/mL GO组、1.0 μg/mL GO组、5.0 μg/mL GO组，采用划痕试验检测划痕后24、36 h HSF的迁移率（样本数为5）及划痕后12 h HUVEC的迁移率（样本数为3），采用酶联免疫吸附测定法检测培养4、6、8 h后HSF分泌的血管内皮生长因子（VEGF）水平（样本数为3）。将配制的含相应终质量浓度GO的GO-GelMA复合水凝胶设为0 μg/mL GO复合水凝胶组、0.1 μg/mL GO复合水凝胶组、1.0 μg/mL GO复合水凝胶组、5.0 μg/mL GO复合水凝胶组，观察其交联前后的性状，检测用磷酸盐缓冲液浸泡3、7 d后GO的释放情况（样本数为3）。在16只6周龄雌性C57BL/6小鼠背部制作全层皮肤缺损创面，将采用原位交联的含相应终质量浓度GO的GO-GelMA复合水凝胶处理的小鼠按随机数字表法分为0 μg/mL GO复合水凝胶组、0.1 μg/mL GO复合水凝胶组、1.0 μg/mL GO复合水凝胶组、5.0 μg/mL GO复合水凝胶组，每组4只，观察治疗3、7、14 d创面大体情况并计算创面愈合率，采用激光多普勒血流仪检测治疗3、7、14 d创面血流灌注并计算平均灌注单位（MPU）比值，采用苏木精-伊红染色观察治疗7 d创面血管新生情况并计算血管密度（样本数均为3）。取0 μg/mL GO复合水凝胶组和0.1 μg/mL GO复合水凝胶组治疗7 d的创面组织，采用苏木精-伊红染色观察GO分布与血管新生的关系（样本数为3），行免疫组织化学染色后观察VEGF的表达。对数据行重复测量方差分析、单因素方差分析、Tukey法。
GO为多层片状结构，宽度约为20 μm、长度约为50 μm。培养48 h，10.0 μg/mL GO组HSF的吸光度值明显低于0 μg/mL GO组（
GO质量浓度低于10.0 μg/mL对HSF增殖活性无明显影响，0.1 μg/mL的GO能够促进HSF和HUVEC迁移，能促进HSF分泌VEGF。原位光聚合GO-GelMA复合水凝胶敷料能够通过促进小鼠全层皮肤缺损创面血管新生，增加创面早期血流灌注，且GO对新生血管有富集作用，其机制可能与GO促进创面细胞分泌VEGF相关。
基于单细胞RNA测序探讨小鼠全层皮肤缺损创面中真皮成纤维细胞（dFb）的异质性与生长因子调控网络。
采用实验研究方法。取5只8周龄雄性健康C57BL/6小鼠（鼠龄、性别、品系下同）正常皮肤组织，另取5只背部全层皮肤缺损小鼠伤后7 d创面组织，用胶原酶D和DNA酶Ⅰ消化组织获得细胞悬液，用10x Genomics平台构建测序文库，用Illumina Novaseq6000测序仪进行单细胞RNA测序。采用R4.1.1软件的Seurat 3.0程序分析获得2种组织细胞的基因表达矩阵，采用按细胞群、细胞来源、标记皮肤中主要细胞基因分类的二维tSNE云图进行可视化展示。根据已有文献报道和CellMarker数据库检索情况，分析2种组织细胞的基因表达矩阵中标志基因表达情况，对各细胞群进行编号和定义。将基因表达矩阵和细胞分群信息导入R4.1.1软件的CellChat 1.1.3程序，分析2种组织中细胞间通信以及创面组织血管内皮生长因子（VEGF）、血小板衍生生长因子（PDGF）、表皮生长因子（EGF）、成纤维细胞生长因子（FGF）信号通路中的细胞间通信，2种组织中FGF的各亚型与FGF受体（FGFR）各亚型的两两配对（以下简称FGF配受体对）对FGF信号网络的相对贡献，2种组织中相对贡献排名前2的FGF配受体对信号通路中的细胞间通信。取1只健康小鼠正常皮肤组织和1只全层皮肤缺损小鼠伤后7 d创面组织，行多重免疫荧光染色，检测FGF7蛋白的表达与分布及其与二肽基肽酶4（DPP4）、干细胞抗原1（SCA1）、平滑肌肌动蛋白（SMA）和PDGF受体α（PDGFRα）的蛋白共定位表达。
健康小鼠正常皮肤组织和全层皮肤缺损小鼠伤后7 d创面组织中均包含25个细胞群，但2种组织中各细胞群中细胞数不一致。
小鼠全层皮肤缺损创面愈合过程中的dFb存在高异质性，为多种生长因子潜在的主要分泌或接收细胞群落，与生长因子信号通路之间存在紧密、复杂的联系；FGF7-FGFR1信号通路是创面愈合过程中的主要FGF信号通路，靶向调控多个dFb亚群。
探讨负载银和重组人碱性成纤维细胞生长因子（rh-bFGF）的甲基丙烯酸酐化明胶（GelMA）水凝胶对兔深Ⅱ度烧伤创面的影响。
采用实验研究方法。制备含不同浓度甲基丙烯酸酐（MA）的低浓度MA明胶（GelMA）材料、中浓度GelMA材料和高浓度GelMA材料，加入光引发剂后分别制得低浓度GelMA水凝胶、中浓度GelMA水凝胶和高浓度GelMA水凝胶。采用核磁共振波谱仪检测前述3种浓度GelMA材料的氢核磁共振谱并根据波谱图计算其取代度，采用场发射扫描电子显微镜（FESEM）检测前述3种浓度GelMA水凝胶的三维微观结构及孔径，样本数均为9。根据前述筛选出的MA浓度合成含10种浓度银的GelMA（含银GelMA）溶液，将每种浓度的含银GelMA溶液均分为3份，加入光引发剂后分别暴露于紫外光下持续20、25、35 s，制得相应的含银GelMA水凝胶。采用胶原酶降解法测定不同光交联时间含银GelMA水凝胶降解12、24、36、48 h的降解剩余率及彻底降解所需时长，样本数为5。测定前述筛选出光交联时间下含10种浓度银GelMA水凝胶对金黄色葡萄球菌的抑菌圈直径反映其抑菌能力，样本数均为5。以与含最低浓度银（即不含银）GelMA水凝胶抑菌圈直径相比有统计学意义的含银GelMA水凝胶为有抑菌活性。选取具有抑菌活性的且载药浓度最低的含银GelMA水凝胶，采用FESEM检测其三维微观结构及孔径，采用能谱仪检测其内部银元素的存在情况，样本数均为9。将冻干单纯GelMA水凝胶和冻干含银GelMA水凝胶分别浸没于磷酸盐缓冲液中24 h，通过称重法计算并比较2种水凝胶的溶胀率，样本数为5。根据预实验及前述实验结果，制备含银和rh-bFGF的GelMA水凝胶（简称复合水凝胶）。大体观察复合水凝胶的外观，并采用FESEM检测其三维微观结构与孔径。取30只4~6个月龄、雌雄各半日本大耳兔，在其背部制作深Ⅱ度烧伤创面。以兔头侧为基准，将脊柱左侧创面作为复合水凝胶治疗组，右侧作为纱布对照组，2组创面分别作相应处理。观察伤后3、7、14、21、28 d创面愈合情况；记录伤后7、14、21、28 d创面愈合面积并计算其愈合率，样本数为30。对数据行重复测量方差分析、单因素方差分析、独立样本
低浓度GelMA材料、中浓度GelMA材料及高浓度GelMA材料的取代度，差异明显（
中浓度GelMA水凝胶在溶胀性、可降解性方面具有良好的理化特性，筛选出的含银GelMA水凝胶具有抑菌活性且载药浓度最低，制得的复合水凝胶可明显缩短兔深Ⅱ度烧伤创面愈合时间。
探讨外源性左旋肉碱对过度内质网应激介导的严重烫伤大鼠肝细胞焦亡的影响及其分子机制。
采用实验研究方法。将15只6~8周龄雌性SD大鼠按随机数字表法（分组方法下同）分为假伤组、单纯烫伤组、烫伤+肉碱组，每组5只，单纯烫伤组和烫伤+肉碱组大鼠背部制作30%体表总面积的Ⅲ度烫伤，其中烫伤+肉碱组大鼠另外给予腹腔注射左旋肉碱。伤后72 h，采用全自动生化仪检测肝损伤生物化学指标天冬氨酸转氨酶（AST）和丙氨酸转氨酶（ALT）水平，样本数为5。伤后72 h，采用苏木精-伊红染色观察肝组织损伤情况。伤后72 h，采用实时荧光定量反转录PCR（RT-qPCR）法检测肝组织中细胞焦亡相关标志物核苷酸结合寡聚化结构域蛋白样受体热蛋白结构域相关蛋白3（NLRP3）、胱天蛋白酶1（caspase-1）、消皮素D和白细胞介素1β（IL-1β）以及内质网应激相关标志物葡萄糖调节蛋白78（GRP78）、CCAAT/增强子结合蛋白同源蛋白（CHOP）的mRNA水平；采用蛋白质印迹法检测肝组织中GRP78、CHOP、NLRP3、caspase-1、caspase-1/p20、消皮素D-N、剪切型IL-1β蛋白表达水平，样本数均为5。取人肝癌细胞HepG2，分为二甲基亚砜（DMSO）组、0.1 μmol/L衣霉素组、0.2 μmol/L衣霉素组、0.4 μmol/L衣霉素组、0.8 μmol/L衣霉素组，分别作相应处理。培养24 h后，采用细胞计数试剂盒8法检测细胞活力并筛选衣霉素干预浓度（样本数为5）。取HepG2人肝癌细胞，分为DMSO组、单纯衣霉素组和衣霉素+肉碱组，分别作相应处理。培养24 h后，采用蛋白质印迹法检测细胞中GRP78、CHOP、NLRP3、caspase-1、caspase-1/p20、消皮素D-N、剪切型IL-1β蛋白表达水平，样本数均为3。对数据行单因素方差分析和LSD-
伤后72 h，单纯烫伤组大鼠血清中AST和ALT水平分别为（640±22）、（157±8）U/L，均分别明显高于假伤组的（106±13）、（42±6）U/L（
在严重烫伤大鼠中，外源性左旋肉碱可能通过抑制过度内质网应激介导的细胞焦亡相关通路发挥对肝损伤的保护作用。
智能驾驶场景下的人车冲突问题与行人过街行为密切相关，为使高级驾驶辅助系统（advanced driving assistance system, ADAS）具备识别行人过街意图的功能，并对人车碰撞事件预警，提出一种基于图表示学习（graph representation learning, GRL）方法的行人过街意图识别框架。它采用开源工具对行人骨架信息进行识别，采用图方法，以行人在一段运动过程内每一帧的骨架关键点为节点，以骨架自然连接关系、相关关系和时域关系为边建立图模型，实现对行人动作序列的表征。以图结构数据为输入，基于支持向量机（support vector machine, SVM）训练行人过街意图识别模型。在自动驾驶数据集PIE上对所提出方法进行评估，结果显示，行人过街意图分类准确率可达90.29%，所提出方法能够有效识别行人过街意图，对提高智能车决策安全性具有重要意义。
根据无人车动态实时避障的需求，提出一种基于人工势场法的局部避障路径规划算法，通过改进势场环境及势场力来解决传统势场法局部极小值和目标不可达的问题. 考虑车辆碰撞安全性，对侧向动态障碍物和同向动态障碍物工况进行分析，采用动态窗口法进行实时动态避障规划. 同时为保证规划路径的平滑性和可跟踪性，采用贝塞尔曲线对轨迹进行平滑处理. 最后,在CarSim和Matlab/Simulink 联合仿真平台下，对所提出的控制算法进行验证. 仿真结果表明了规划算法的避障有效性、安全性以及可跟踪性.
智能车辆轨迹跟踪的准确性与鲁棒性是车辆运动控制性能的重要表征，基于路径预瞄信息的跟踪控制研究使车辆性能显著提升. 然而，车辆转向系统响应不足给车辆实时准确的基于预瞄信息跟踪参考轨迹带来挑战. 针对此问题，实时引入转向系统状态建立双闭环轨迹跟踪控制结构，保证智能车辆轨迹跟踪控制算法对转向系统响应不足的鲁棒性. 具体结构外环基于预瞄信息使用模型预测控制求解最优转向角，内环基于转向状态误差使用PID方法设计反馈控制律以补偿转向响应不足. 双闭环结构耦合控制输入保证了车辆鲁棒最优跟踪控制. 最后通过Carsim与Simulink联合仿真，验证了该双闭环控制结构的有效性.
车辆质心侧偏角对于车辆横向稳定状态判断具有重要作用，对质心侧偏角的高估或低估都会对稳定性控制系统产生影响. 针对目前质心侧偏角估计方法仍具有较大误差且实用性不强，提出了以降低观测误差及提高估计系统实用性为目标的方法，构建了鲁棒性较强的模糊二阶滑模观测器计算质心侧偏角观测值，同时采用惯性测量单元信号计算质心侧偏角积分值. 之后分析了两种估计方法的优缺点，对质心侧偏角观测估计值与传感器信号积分估计值进行可拓融合，以实现采用传感器信号估计对观测值进行修正. 最后通过Simulink/TruckSim仿真、硬件在环仿真，进行了质心侧偏角估计方法的验证. 在实车定圆加速测试工况中以控制效果论证了所提出方法的有效性. 研究表明所提出方法能够准确反映实际质心侧偏角状态，并且可靠性、实用性均较佳.
考虑转向控制系统故障和未知干扰同时作用对无人车辆路径跟踪效果的影响，为提高无人车辆控制系统的可靠性，设计了一种无人车路径跟踪容错控制方法. 对转向控制系统输入性故障进行分析，结合未知干扰情况定义了名义故障并建立相应的数学模型. 利用高阶滑模观测器构造名义故障微分方程，并利用自适应容积卡尔曼滤波设计了车辆质心侧偏角和名义故障估计方法，从而为无人车容错控制提供可靠信息源. 基于滑模控制理论设计了无人车路径跟踪容错控制器并证明了其收敛性. 联合仿真和硬件在环试验结果表明，所提出的估计方法能够得到精确可靠的质心侧偏角和名义故障估计结果，且与无容错控制相比，所设计的路径跟踪容错控制器在面对故障和干扰时能够明显地提高车辆的控制性能，并同时保证车辆的路径跟踪能力及其自身的稳定性.
智能车辆相关技术已实现了长足的发展，并已能够在有限封闭场景中实现自主行驶的基本功能. 然而，实际道路测试结果表明，目前智能车辆技术仍存在较多局限，而智能车辆在复杂城市与越野环境的大规模应用仍面临较多挑战. 作为智能车辆关键技术之一，运动规划与控制技术已基本建立了完整的理论体系并已得到较多工程应用，但传统方法在实际应用中仍存在动态复杂场景理解能力弱、场景适应性差、模型复杂度高、参数调整难度大等缺陷. 由于机器学习方法具备较强的知识表征与模型拟合能力，其已经在智能车辆的感知与导航技术中得到了广泛的应用. 而为了解决传统运动规划与控制技术存在的泛化性与适用性等问题，许多研究者近年来也开始探索基于深度学习、强化学习等机器学习方法的运动规划与控制方法. 本文将对目前基于机器学习的智能车辆规划与控制方法研究现状进行回顾，从规划与控制策略基本架构、基本学习范式以及基于学习的规划与控制方法三方面对现有智能车辆规划与控制策略学习方法进行分析，最后对研究现状与未来发展方向进行总结与展望.
基于智能汽车行驶的空间约束和运动轨迹曲率约束等限制，对换道轨迹的燃油经济性进行了研究. 基于发动机瞬态油耗模型，确定了平路行驶的最经济车速，并建立了能满足各种约束的3阶贝塞尔换道轨迹模型. 通过Matlab/Simulink与Carsim联合仿真验证，文中设计的贝塞尔换道轨迹的油耗分别比已有的sin-tanh和
陆空平台具有多域机动能力，通过陆空模式的转换能够适应各种复杂环境，但陆空模式转换多为静止起飞或悬停下降，这种静态起降方式不利于陆空平台机动性的充分发挥. 针对一种动力机构可偏转的变构型陆空两栖平台，基于牛顿-欧拉方程建立陆空平台的飞行动力学模型，规划偏转角的时间序列以获得动态动力学约束，确定相对时间最优目标函数；基于5次多项式拟合二维平面轨迹，根据PID控制方法设计轨迹跟踪控制器，并进行轨迹规划和控制仿真. 结果表明，动态切换时间相比静态切换时间缩短了23.02%，动态切换规划轨迹平滑，高度方向无超调，控制器能较好地跟踪目标飞行轨迹.
为定量描述胶结充填体在动载作用下的损伤程度及破坏过程，利用数值模拟软件对胶结充填体进行SHPB动态冲击，并通过室内SHPB冲击试验结果验证数值模拟方法的可行性. 对不同冲击速度（1.5 ，1.7 ，1.8 ，2.0 m/s）条件下4种配比胶结充填体（灰砂质量比分别为1∶4, 1∶6, 1∶8, 1∶10），采用微裂纹密度法定义损伤变量值
为解决同步磁阻电机转子结构复杂、机械强度差、难以应用于高速领域的缺点，提出了一种新型同步开关磁阻电机（synchronous switched reluctance motor，SSRM）. 该电机结合了开关磁阻电机与同步磁阻电机的优势，电机转子冲片制作工艺简单、可靠性强、适用于高速领域，电机的凸极率高，转矩输出能力强. 在对新型SSRM电磁设计的基础上利用正交法对该电机参数进行优化，选取转矩均值、转矩脉动以及电机效率作为优化性能指标. 通过有限元仿真和实验验证了优化后的SSRM具有转矩脉动低、效率高的特性.
微表情顶点帧蕴含着丰富的微表情信息，为了准确地检测出微表情顶点帧，本文提出了一种基于光流特征的神经网络分类，并利用先验知识规则进行取舍的检测方法. 该方法针对固定滑窗大小内的图像进行光流信息提取，利用双输入特征提取网络对
针对当前关于双隐身飞机编队相干干扰对单脉冲雷达具体影响分析不足及误差有效性缺乏合理评价的问题，考虑双机回波相位差的随机性特点，建立了双隐身飞机相干干扰模型和角度误差评价模型. 设定双机突防运动场景，提取双机动态RCS，推导回波作用下的双机相干干扰引起的随机性角度误差计算式，求解随机性角度误差的一阶数字特征，并根据脱靶距离建立了角度误差有效性影响评价准则，解算角度误差的有效影响概率. 仿真表明：双隐身飞机相干干扰产生的角度误差均值较大，随机起伏性较弱，干扰效果较好，且误差对雷达影响有效概率较大，保证了编队突防安全性.
针对船用钢材料在超高应变率下的动态响应机制及变形强化机理尚不明确的技术基础问题，通过一维平板撞击试验测得了10，20及30 GPa撞击压力下E36船用钢的自由表面速率−时间曲线，计算得到了E36船用钢的Hugoniot弹性极限和层裂强度，利用ANSYS软件模拟了不同撞击压力下的温度场；并采用SEM、TEM等技术研究了E36船用钢在高压撞击下的损伤演化规律和变形强化机理. 试验结果表明：不同撞击压力下材料均发生了层裂，毁伤机理为微孔和微裂纹形核、长大和聚合；随着撞击压力的增加，E36船用钢的Hugoniot 弹性极限变化不大，层裂强度逐渐增加，相变强化、位错强化和孪晶强化是E36船用钢在高压、高应变率下的主要强化机制.
针对冰晶在温暖环境下运动时固液汽耦合的相变现象，基于欧拉法建立了冰晶粒子的运动相变模型和计算方法。计算了冰晶粒子在强迫对流环境下的融化相变过程，与实验结果对比验证了运动相变模型和计算方法的准确性。针对NACA0012翼型计算了冰晶绕流运动时的热力学特性，得到了冰晶粒子到达撞击表面时的融化状态与收集系数。研究了冰晶粒径大小、初始球形度、气流相对湿度和温度对运动相变的影响。结果表明：冰晶粒子运动相变模型可以有效地评估冰晶结冰风险，冰晶粒子的融化速率主要取决于粒子直径、球形度、气流温度、湿度等因素，环境温度为288 K时冰晶粒子的融化时间为27.5 s，而相同条件下环境温度为302 K时的融化时间仅有5.2 s。
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