O-linked-N-acetylglucosaminylation (O-GlcNAcylation) is a type of glycosylation that widely present in eukaryotic cytoplasmic proteins or mitochondria. It affects protein properties, cell functions, disease states and so on, hence it is import to identify the roles that O-GlcNAcylations play on target proteins in living cells. The new technologies for manipulating O-GlcNAcylations on target proteins will greatly accelerate the understanding of O-GlcNAc’s functions. This article briefly introduces some recent research progress of chemical biology techniques for targeted protein O-GlcNAcylation, and also analyzes the advantages and limitations of these strategies, and their future development prospects. These technologies build a powerful chemical biology toolbox, which may contribute to the diagnoses and treatments related to O-GlcNAcylation.

Antibiotic resistance is considered one of the most serious global threats to human health in the 21st century. The overuse of antibiotics has aggravated the development of bacterial resistance. Therefore, it is necessary to further study the mechanisms of bacterial resistance and explore new types of resistance and bacteriostatic strategies. This article provides an overview of the mechanisms of bacterial resistance and new antibacterial strategies for drug-resistant bacteria. It elaborates on the molecular mechanisms of three types of resistance: inherent resistance, acquired resistance and adaptive resistance. It also discusses new antibacterial strategies such as new antibacterial compound molecules, phage therapy, CRISPR-Cas system therapy and antisense therapy. This review aims to summarize the mechanisms of bacterial resistance and provide a reference for subsequent prevention and treatment of bacterial resistance.

The central dogma of molecular biology explains the direction of genetic information flow within organisms, and is one of the most important classical theories in molecular biology. There are two versions of the central dogma. In 1957, F. Crick first proposed the central dogma, which was published in 1958 and revised in 1970. Crick’s central dogma divides genetic information transfer modes into three groups: (l) three types of general transfer that occur normally in cells, including DNA→DNA, DNA→RNA, and RNA→protein; (2) three types of special transfer refer to RNA→RNA and RNA→DNA in certain viruses, as well as in vitro DNA→protein; (3) three types of undetected transfer are information transmissions that have not yet been discovered or may not exist, i.e. protein→protein, protein→DNA, and protein→RNA. In 1965, J. Watson proposed another version, using the protein biosynthesis pathway as the central dogma, which divided the process of genetic information transfer into two steps:transcription and translation, usually simplified as DNA→RNA→protein, and later supplemented with “RNA replication” and “reverse transcription”. The limitations of Watson’s version have been questioned.

Epidermal growth factor receptor (EGFR) is a receptor tyrosine kinase that participates in physiological processes such as cell proliferation, division, and differentiation. EGFR plays an important role in the occurrence and development of tumors. In targeted treatment of non-small cell lung cancer, tyrosine kinase inhibitors (TKIs) targeting EGFR have achieved significant efficacy. However, with the emergence of mutations such as EGFR T790M, patients may develop resistance to the first and second-generation EGFR TKI therapy. Therefore, the third generation EGFR TKI, represented by Osimertinib, has achieved good results in patients with EGFR T790M mutations. However, some patients still develop acquired drug resistance after third generation EGFR TKI treatment. Currently, the drug resistance mechanisms are mainly divided into two types: EGFR dependent (mutations in the EGFR kinase domain) and EGFR independent (activation of abnormal bypass signals and downstream signal pathways, and histological phenotypic changes). This article comprehensively reviews and summarizes the structure and main drug resistance mechanisms of the third generation TKI, and the treatment strategies after drug resistance and the possible directions for overcoming EGFR TKI resistance in the future are also analyzed.

Covalent attachment of glycans to the side chains of protein amino acids can form glycosylation. Glycosylation, in many cases, significantly increases the diversity of protein structure, properties, and functions. Specific glycosylation patterns can confer specific effects on proteins and enzymes, while aberrant glycosylation patterns may lead to diseases. Therefore, a comprehensive understanding of the role of protein glycosylation is of great significance, both for basic and applied research. However, progress in this area has been exceptionally slow due to the difficulty in obtaining suitable samples for study. In recent years, researchers have gradually begun to explore the use of a library-based strategy to change this situation. This strategy involves the synthetic preparation and characterization of a series of homogeneously glycosylated protein isoforms (glycoforms) with high purity and systematic variations in structures. Through the following comparative analysis, the effects and functioning mechanisms of protein glycosylation can be relatively quickly and accurately obtained. This, in turn, enhances the application of glycosylation in improving the performance of proteins and enzymes. This review aims to summarize, outline, and briefly discuss the progress in the O-glycosylation research direction. It presents research methods and achievements of a few existing examples in chronological order, with the aim of helping researchers gain a clearer understanding of the current development and shortcomings of this strategy. This review is expected to assist researchers in better utilizing this strategy for protein glycosylation research and applications, ultimately improving the depth and breadth of this research.

In eukaryotes, mRNA translation includes initiation, extension, and termination. Among them, the initial stage of translation is the most important and complex, which determines whether mRNA can be effectively translated into protein. Eukaryotic mRNA can be divided into traditional cap-dependent translation and alternative cap-independent translation according to translation initiation mechanism. When the stimulation of the external environment or the changes of the cell itself make the cell in a state of stress, the traditional cap-dependent translation is inhibited or downregulated, and the alternative mechanism of cap-independent translation is activated to ensure the smooth progress of translation. In order to maintain the demand of protein synthesis under stress, eukaryotes employ various translation initiation mechanisms to achieve cap-independent translation. The more common are translations initiated by internal ribosome entry site (IRES), m⁶A modifications, and cap-independent translation enhancer (CITE). These mechanisms allow ribosomes to initiate within specific regions of mRNA without relying on the traditional m⁷G cap structure. Through these mechanisms, eukaryotes can still synthesize proteins to meet the needs of cells under stress.Based on a summary of the traditional cap-dependent translation research, this paper focuses on the cap-independent translation mechanism initiated by IRES, m⁶A and CITE, in order to provide references for better exploration of the translation mechanism under cellular stress, and provide more ideas for future translation research and application.

Glucose is the most important biological energy donor, is the source of energy and metabolism of living cells, and plays an important role in cell signaling. Abnormal glucose metabolism is frequently associated with diabetes mellitus, retinopathy, Alzheimer's disease, and cardiovascular diseases. Sodium-glucose cotransporter 2 (SGLT2) is one of the transporters involved in the maintenance of glucose homeostasis. It mediates the reabsorption of glucose by the kidney and plays an important role in maintaining the stability of plasma glucose. Sodium-glucose cotransporter 2 inhibitors (SGLT2i) can reduce the reabsorption of glucose in the kidney and have a positive impact on blood glucose, blood pressure, body weight, etc. This paper summarizes the current research progress of SGLT2 and its inhibitors, the structure and key sites of SGLT2 and the mechanism of SGLT2i. In particular, in recent years, it is found that the protective effect of SGLT2i on kidney and cardiovascular is more obvious, and it can also resist the occurrence and development of tumor cells. Finally, this paper summarises and discusses the current bottlenecks in research related to SGLT2i and provides new ideas for future structure-based drug optimisation of inhibitors.

Hematopoietic prostaglandin D synthase (HPGDS) is a glutathione-transferase enzyme that is widely distributed in a range of tissues and is found in megakaryocyte lineages as well as a number of immune cells. HPGDS has been identified to be involved in the regulation of allergic reactions in the body, accelerating the conversion of prostaglandin H2 (PGH2) to prostaglandin D2 (PGD2), and the binding of PGD2 to DP2 receptors increasing the release of inflammatory cytokines and the development of allergic reactions. In recent years, it has been discovered that HPGDS has a substantial role in allergic rhinitis, allergic asthma, eosinophilic esophagitis (EoE), and atopic dermatitis (AD). With Th2 cells in atopic dermatitis, mast cells in allergic asthma, eosinophilic granulocytes in allergic rhinitis and eosinophilic esophagitis, and other key players in various diseases, HPGDS plays a critical role in the pathogenesis of allergic reactions and inflammatory diseases and is essential for disease development. Notably, medications that target HPGDS such as HQL-79, TAS-204, TAS-205, and TFC-007 can effectively treat the symptoms of many allergy disorders in which HPGDS is excessive. HPGDS can be a significant target for the treatment of associated diseases. In order to highlight the significance of HPGDS in the pathogenesis of allergic diseases and to offer fresh ideas for the creation of medications for the treatment of allergic diseases, this paper describes the biological functions of HPGDS, reviews the important role and research progress of HPGDS in allergic diseases, as well as the experiments of HPGDS-related target drugs.

O-linked β-N-acetylglucosamine (O-GlcNAc) modification is a widespread post-translational modification of intracellular proteins. Unlike common types of protein glycosylation, O-GlcNAc transferase (OGT) adds a single GlcNAc unit to serine or threonine residue of proteins. Since its discovery, a large number of studies have shown that O-GlcNAcylation is widely involved in many fundamental physiological processes such as cell growth and development, gene transcription, immune response and stress response. In the immune system, O-GlcNAcylation regulates the activation, differentiation and function of immune cells through various ways. The differentiation and phenotypic maintenance of macrophages are dependent on O-GlcNAcylation, the fluctuation of glucose metabolism levels or the loss of OGT will lead to the transformation of macrophage polarization. In addition, O-GlcNAcylation regulates cytokines transcription by altering the activity of transcription factors such as NF-κB to maintain macrophage inflammation, and also affects MAVS (mitochondrial antiviral signaling) protein ubiquitination in response to pathogen infection. In other innate immune cells, the reduction of O-GlcNAcylation will affect cellular immune function to varying degrees. O-GlcNAc regulates transcription factors such as NF-κB, NFAT (nuclear factor of activated T cells) and c-Myc in T cells and B cells, affects the expression of cytokines and metabolism-related genes, and meet cell activation and proliferation by a higher level of glucose intake. The abnormal changes of O-GlcNAcylation in the immune system are closely related to the occurrence and development of chronic inflammation, tumor and other related diseases, or become a means of tumor immune escape. An in-depth understanding of the role of O-GlcNAc modification in the immune system will help to reveal the molecular mechanism of immune regulation and provide a theoretical basis for the development of novel immunotherapy strategies.

The nucleolar protein N-acetyltransferase 10 (NAT10) is responsible for catalyzing the acetylation of both lysine and cytidine residues. At present, plenty of studies have revealed that these acetylation modifications play vital roles in numerous physiological processes, such as telomerase activity, stress response, DNA damage repair, cell cycle regulation, rRNA biosynthesis regulation, mRNA stability, and translation. Additionally, NAT10 has a strong association with the progression and prognosis of human cancers and Hutchinson-Gilford premature aging syndrome (HGPS). However, there are still some limitations. For example, the complete structure of NAT10 and the impact of these structures on functions are still unknown, the cellular functions mediated by NAT10 remain unclear and the exact mechanism by which NAT10 influences human cancers and HGPS progression is needed to be clarified. This review covers the structure, enzymatic activity, biological functions, as well as roles in diseases of NAT10. We also propose the limitations of current research and envision the future research directions, aiming to provide a reference for NAT10 research.

糖作为生命体的重要组成成份,承载着很多生物学功能:保护细胞、细胞的结构成份、能量来源与代谢组分、细胞与细胞的识别以及细胞内的信号转导。最常见的人类ABO血型的分子基础就是由血细胞表面的糖链结构决定的,流感病毒侵入宿主细胞则是由宿主细胞表面的唾液酸糖作为受体。
虽然人们很早就认识到了糖的重要性,但其固有的复杂性造成了很多研究上的技术难题:其一,化学结构上,糖链的链接方式多种多样;其二,糖链的合成没有模板,而是如同“分子乐高”一样由糖基转移酶装配合成。这样的糖类异质性虽然可以承载更多的信息,但是也为研究它的生物学功能造成了不小的挑战。
所以糖生物学从诞生的那一刻起,就注定了是一门多学科多手段多角度的交叉学科。本专刊涵盖多学科(生命、化学以及药学)对于糖生物学功能的综述以及技术上的最新简报,力图报道糖科学研究的新技术新方法,阐释糖在不同生理和病理中的功能,并讨论糖类药物的开发。
本专刊一共收录了9篇糖生物学相关的综述以及1篇技术简报。
(1) “工欲善其事,必先利其器”,我们首先综述了研究糖生物学的技术手段:《基于分子库策略的蛋白质O-糖基化研究》通过化学合成的方法来研究不同Ser/Thr位点进行O-糖基化对蛋白质功能的影响和调控;《酶介导的邻近细胞标记方法探究细胞间相互作用》阐释了化学生物学的研究策略;《基因编辑技术及其在糖生物学研究中的应用》则将糖与基因编辑相结合,对未来进行了展望。
(2) 聚焦N-乙酰葡萄糖胺 (O-linked β- N -acetylglucosamine, O-GlcNAc)。O-GlcNAc为发生在细胞内参与信号转导的单糖修饰。综述《靶向编辑O-GlcNAc糖基化修饰的化学生物学技术》从化学的角度阐释了最新的化学生物学工具;技术简报《CpOGA^D298N与核心链霉亲和素(Stv13)融合表达用于检测蛋白O-GlcNAc修饰》则从去N-乙酰葡萄糖胺修饰的角度研究了新的检测方法;《O-连接-N-乙酰葡糖胺糖基化蛋白质的富集方法》介绍了近年来 O-GlcNAc 糖基化修饰与疾病之间的关系以及相关修饰位点的富集方法;《O-GlcNAcylation在免疫系统中的作用》则侧重其在生物学中的免疫功能。
(3) 阐释糖类分子在不同病理中的功能:《免疫分子的糖基化修饰与重要感染性疾病》集中于免疫系统;《溶酶体半乳糖苷酯酶作用机制及疾病》侧重糖在溶酶体这一独特的细胞器中的功能。以此为基础,《糖药物在疾病治疗中的应用》综述了针对糖类的药学研究。
我们相信通过多学科对于糖生物学的交叉探索与创新实践,糖生物学必将迎来崛起的新时代。

To adapt to the burgeoning diversity and complexity of new viruses, existing virus classification methods have revealed their limitations. Addressing these challenges, the International Committee on Taxonomy of Viruses (ICTV) has rolled out a novel classification system in 2019, comprising 15 taxonomic ranks or taxa. This system, available on the ICTV's official site: https://ictv.global/, allows the public to freely access real-time virus taxonomy data. This system sheds light on the intricate connections and macro evolutionary mechanisms of viruses, allowing a more objective and systematic approach for virus classification and nomenclature.

Carbohydrates play a crucial role in various life processes such as cell recognition, bacterial infection, signal transduction, and immune response. Due to their associated biological effects, the naturally existing carbohydrates and their derivatives have been extensively studied. Carbohydrates are also important lead for drug development, carbohydrate-based drugs exhibit excellent therapeutic efficacy in the treatment of various diseases such as infection, cancer, and cardiovascular disease. Ribose and deoxyribose are the primary scaffolds for nucleoside drugs, which can inhibit viral replication and serve as treatments for viral infections. Carbohydrates-containing macrolide and aminoglycoside antibiotics target the 50S subunit and 30S subunit of bacterial ribosomes respectively, to hinder protein synthesis and eliminate bacteria. Heparin, a highly sulfated glycosaminoglycan, acts as an anticoagulant by binding to antithrombin III and inactivating thrombin. Pseudo-saccharide can bind to glycosidase to prevent oligosaccharide hydrolysis, thereby controlling blood sugar levels. Additionally, sugar vaccines are crucial in cancer treatment, highlighting the broad applications of sugar drugs across various diseases. Furthermore, the rapid development of glycochemistry has deepened scientists’ understanding of carbohydrates, and medicinal chemists increasingly apply this knowledge to the design of new drugs. This review provides a brief overview of the application of carbohydrate-containing drugs in various disease.

The development of the skull depends on the normal bone supply of the cranial suture. During this period, the cranial suture mesenchymal stem cells undergo systematic proliferation and osteogenic differentiation to ensure the normal growth of the bone. Cranial suture cells, extracellular matrix, periosteum and meninges form the cranial suture niche and regulate each other to ensure the normal development of the cranial suture. Clinical studies have found that the vast majority of patients with craniosynostosis are accompanied by abnormalities of dura mater and extracellular matrix (ECM). Many studies have also found that abnormal external mechanical stimuli acting on cranial sutures may also lead to abnormal cranial suture development. Considering that the development of cranial sutures is affected by many factors, this review will introduce the research progress of microenvironment in abnormal cranial suture and mechanical force in the regulation of cranial suture development in recent years, and summarize the molecular mechanism and current research direction of craniosynostosis, so as to provide new ideas for the diagnosis and treatment of the disease.

With the rapid development of cell therapy, large-scale lentiviral production has become a bottleneck in the process chain; thus, optimizing the production process of CAR lentiviral vectors with high titer and high purity of 293T becomes crucial. This study aimed to optimize the packaging of lentiviral 293T adherent cells to achieve time-saving, cost-saving, and improved lentiviral packaging. At the same time, the optimization of lentiviral vectors was carried out to explore the factors affecting the growth of suspension cell clusters. The 293T adherent cells were domesticated into suspension culture by fast and slow domestication. Their cell morphology, cell density, cell viability, lentiviral packaging ability, and stable consistency after cryopreservation and resuscitation were compared to screen out the optimal suspension domestication conditions. Comparative cell clumping growth was studied by adjusting Ca²⁺ concentration and EDTA addition. The findings showed that the serum-free medium OPM-293 CD05 Medium could be quickly domesticated into 293T suspension cells from 293T adherent cells, and that these suspension cells could be produced with lentiviral titers that were better than the adherent cells’ packaging titers (^*P<0.05). Ca²⁺ concentration affects the size of cell clusters. The addition of EDTA effectively separates and disperses unnecessary cell clusters. In summary, the experiment’s findings demonstrated that serum-free OPM-293 CD05 Medium could quickly domesticate conventional 293T adherent cells into suspension cells. Within a certain range, the higher the concentration of Ca²⁺, the larger the agglomerates and particle size, and the higher the addition of EDTA, the smaller the agglomerates and particle size.This provides a theoretical framework for the optimization of suspension culture conditions and the lentiviral vector packaging process. It also establishes a theoretical framework for the scale-up and manufacturing of in vitro cell culture, which has some practical applications.

Alzheimer’s disease is a common neurodegenerative disease that seriously affects the quality of life of patients. At present, there is no effective targeted treatment. The pathogenesis of AD is complex and results from the combined effects of environmental, genetic and age factors. The typical pathological features of AD are the deposition of β-amyloid (Aβ), neurofibrillary tangles formed by hyperphosphorylation of microtubule-associated protein tau, and neuronal loss. A large number of studies have demonstrated that the aggregation of Aβ and tau proteins induces nucleotide-binding oligomerization domain-like receptors containing Pyrin-domain protein-3 (NLRP3) inflammasome activation, which is the core of the inflammatory mechanism of AD. Moreover, inhibitors and compounds targeting NLRP3 inflammasome and its upstream and downstream molecules can play a neuroprotective role in cells and animal models and improve spatial memory dysfunction, but clinical efficacy and safety remain to be studied. Therefore, inhibition of NLRP3 inflammasome activation may be a potential therapeutic target for AD. In this paper, the activation mechanism and influencing factors of NLRP3 inflammasome as well as its relationship with AD are reviewed, and the therapeutic drugs targeting NLRP3 inflammasome for AD are summarized in order to provide a new direction for NLRP3 inflammasome related diseases such as AD.

The skin, being a highly intricate organ within the human body, has long posed a challenging and significant aspect in both clinical and research domains when constructing a bionic skin model. 3D bioprinting technology has effectively addressed this issue by facilitating the creation of skin constructs. This innovative technique allows for the rapid and dependable production of bionic skin substitutes, as it enables the customization of skin shapes and the precise distribution of cells and other materials to meet the specific demands of clinical and industrial applications. Furthermore, this technology exhibits exceptional performance, adaptability, superior resolution, reproducibility, and high throughput, rendering it suitable for diverse applications such as drug screening, cosmetic testing, and skin grafts. Consequently, it holds the potential to serve as a viable skin substitute for a larger population of individuals afflicted with burns or diabetes. The integration of 3D bioprinting technology with stem cells has provided researchers and medical professionals with increased opportunities to address skin diseases. Pluripotent stem cells are particularly promising due to their ethical acceptability and ability to be genetically customized to meet individual patient’s needs. This customization enables the development of cell carriers with predetermined structures, which can incorporate biomimetic hierarchies, including blood vessel networks, hair follicles, and sweat glands. These advancements aim to enhance tissue functionality and improve the aesthetic outcomes of implantation procedures. This article summarizes the current research progress in skin construction using 3D bioprinting technology. It specifically highlights the distinctive features of skin construction achieved through various 3D bioprinting techniques. Additionally, it analyzes the specific requirements for skin construction that includes appendages, aiming to offer valuable insights for the development of in vitro physiological or pathological models, regenerative medicine, and clinical research within this domain.

The elucidation of gene functions is a fundamental task in modern biological research, the precisive, efficient, and targeted editing of genes is an indispensable tool for their functional dissection. Over the past 30 years, gene editing has experienced the transformation from homologous recombination repair-based technology to programable nuclease-based method, such as zinc-finger nucleases, transcription activator-like effector nucleases, and CRISPR-associated nucleases. The development of these technologies has greatly advanced the investigation of gene functions and led to the emergence of disruptive technologies for disease treatment. In this manuscript, we first introduced the classification, composition, and working principle of CRISPR-Cas for the immune defense in bacteria. We subsequently focus on the state-of-the-art gene editing tools based on the CRISPR-Cas system including the CRISPRi/CRISPRa, base editors, precursor editors, and the RNA-targeted RCas editing system. We then review the strategies for the delivery of gene editors to the desired target cells/organs, we mainly discuss the pros and cons of adeno-associated viruses, lipid nanoparticles, and extracellular vesicles. Finally, we review the applications of gene editing technologies in glycobiology research, including the function, biosynthesis, and underlying mechanism for the carbohydrates, glycoproteomic analysis, the construction and application of cellular glycan array, and the protein glycoengineering. In conclusion, the development of the precise gene editing technology has significantly promoted the research on the biosynthesis, structure, and function of carbohydrates, which has also advanced the translational aspect of glycoscience research.

Hydrogen sulfide (H₂S) is the third endogenous gasotransmitter, following nitric oxide and carbon monoxide, which regulates various physiological and pathological processes by influencing cellular signaling pathways. In recent years, research has found that the effects of H₂S are mainly mediated through sulfhydration of target proteins, which can alter their structure, affect their activity, stability and protein-protein interactions, thus regulating cellular signaling pathways and related biological processes.This article reviews the current research of sulfhydration and provides a detailed summary of the known sulfhydration modifications and their specific cysteine (Cys) sites that have been identified. This article further discusses the relationship between sulfhydration and nitrosylation.It also covers the types of reactions involved in sulfhydration modification and describes the principles and characteristics of three chemical detection methods: the maleimide assay, the modified biotin switch assay, and the tag-switch assay. By comparing the conditions and applications of sulfhydration detection using these three methods, this article further summarizes the important procedures during the detection process.Based on our experience in sulfhydration research, this article focuses mainly on the cell and tissue sample preparation with detailed experimental procedures for the maleimide assay. Additionally, it discusses the limitations of current sulfhydration detection methods and the unresolved issues in this field, which may provide references for further research in sulfhydration.

Endoplasmic reticulum stress (ERS) is a protective cellular response that occurs when cells face hypoxia or nutrient deprivation. It alleviates protein accumulation in the endoplasmic reticulum (ER) by the unfolded protein response. Unfolded or misfolded proteins that are not efficiently cleared by the unfolded protein response pathway are degraded by endoplasmic reticulum-phagy (ER-phagy), which is trigged by ERS to restore ER morphology. ER-phagy is mediated by ER-phagy receptors. In mammalian and yeast cells, various ER-phagy receptors exist that promote ER fragment formation, capture autophagic cargos and deliver them to autolysosomes for degradation. Each ER-phagy receptor has unique structural features that determine its mode of cargo capture. Additionally, ERS regulates ER-phagy by mediating the expression and phosphorylation of ER-phagy receptors. Research has shown that ERS-induced ER-phagy plays a crucial role in the pathogenesis of various human diseases. Therefore, elucidating the specific mechanisms underlying ERS-induced ER-phagy provides a theoretical basis for the prevention and treatment of ER-phagy-related diseases. Herein, we review the molecular mechanisms of ERS-induced ER-phagy mediated by ER-phagy receptors in mammals (FAM134B, RTN3L, SEC62, CCPG1) and yeasts (Atg39, Atg40, Erp1), as well as the connection between ERS-induced ER-phagy and human diseases such as neurodegenerative disorders and cancer, aiming to provide new strategies for the prevention and treatment of ER-phagy-related diseases.

Colorectal cancer ranks among the malignancies with high incidence and mortality rates, posing significant challenges to its prevention and treatment. In recent years, accumulating evidence has highlighted the critical involvement of deubiquitinases in the development and progression of colorectal cancer. Deubiquitinases meticulously remove ubiquitin moieties from proteins, thereby regulating protein stability, cellular signaling cascades, and gene expression, which in turn impacts key processes in tumor cells such as proliferation, survival, and metastasis. Deubiquitinases can influence the stability of cell cycle proteins, promoting cell cycle progression and accelerating cellular proliferation. Within the Wnt/β-catenin signaling pathway, deubiquitinases contribute to pathway hyperactivation by enhancing nuclear localization of β-catenin, a pivotal event in colorectal cancer initiation. Deubiquitinases also play a role in modulating the stability of immune checkpoint regulators, affecting the function of immune cells within the tumor microenvironment and facilitating immune evasion. Through regulation of transcription factor ubiquitination status, deubiquitinases impact target gene expression, promoting epithelial-mesenchymal transition, thereby augmenting colorectal cancer's invasive and metastatic potential. Moreover, deubiquitinases mediate chemoresistance in tumor cells by controlling the stability of apoptosis inhibitors, DNA repair enzymes, or drug efflux pumps.Given the critical role of deubiquitinases in colorectal cancer progression, the development of small molecule inhibitors targeting specific deubiquitinases has emerged as an attractive yet challenging field of research. Several inhibitors have demonstrated the capability to inhibit colorectal cancer cell growth and induce apoptosis in vitro and animal models. This review delves into the advancements in understanding the roles of deubiquitinases in colorectal cancer and discusses the application of small molecule inhibitors in colorectal cancer, providing insights for therapeutic strategies against this disease.

Cell-cell interactions (CCIs) can occur through the formation of intercellular synapses mediated by cell surface proteins, glycans, lipids, to maintain body homeostasis and regulate physiological functions.These CCIs are complex, involving the participation of many different cell surface and intracellular molecules. Therefore it is key to accurately identify, characterize and quantify cell-cell interactions. In recent years, the technical means of researching CCIs have been continuously introduced, among which proximity labeling is a promising chemical biology method for studying cell-cell interactions.Currently, there are mainly two types of labeling strategies. One is to rely on the direct binding between enzymes expressed on the surface of “bait” cells by genetic engineering and receptor substrates on adjacent cells to achieve intercellular proximity labeling. The other is to use enzymes or organocatalysts (such as photocatalysts) that are recombinantly expressed by genetic engineering or coupled to the surface of “bait” cells by chemical (chemoenzymatic) methods, and following appropriate stimulation or activation, targeted delivery of labeling molecules are carried out. Between them, the enzyme-mediated proximity cell labeling methods have promising application value in the detection and characterization of CCIs.This review defines methods involving enzymes during the labeling process as enzyme-mediatedproximity cell labeling methods. A remarkable advantage of this approach is the small labeling radius that can be achieved due to direct physical contacts between the enzymes and receptor substrates or enzyme-catalyzed generation of highly reactive labeling molecules.We summarize the principles, advantages and disadvantages,and existing applications of the enzyme-mediated proximity cell labeling methods developed in recent years.

Neural tube defects (NTDs) are common, severe, and complicated congenital malformations that are the result of the interaction and interplay of genetic, nutritional, and environmental factors. Maternal periconceptional folic acid (FA) supplementation is a significantly effective strategy for primary prevention of a proportion of NTDs. However, there still exists a portion of NTD cases that are intrinsically resistant to FA therapies, which are widely referred to as “FA-non-responsive NTDs” or “FA-resistant NTDs” . These diseases have a complex etiology, involving genetic, nutritional, environmental and maternal factors. This review integrates the genetic, nutritional, environmental and maternal risk factors associated with FA non-responsive NTDs and reveals the progress of research on their pathogenic mechanisms. Among them, the genetic factors cover three aspects: mouse mutants and strains, FA one-carbon metabolism genes, and key apoptosis genes, which provide possible genetic testing loci for prenatal diagnosis of these children. Nutritional factors concentrate on the roles of inositol and methionine, delineating their possible mechanisms of action and suggesting new directions for early nutritional interventions in NTDs management. In addition, this paper explores the possible mechanisms by which vitamin B12, a cofactor in the FA one-carbon metabolism, in FA-non-responsive NTDs development. We posit that a combination of FA and other vitamins may enhance treatment strategies for these malformations. At last, this paper also reviews the influence of environmental and maternal factors on FA non-responsive NTDs, aiming to provide health recommendations for early pregnancy for at-risk populations. In summary, this article reviews and evaluates research advancement concerning FA non-responsive NTD risk factors and pathogenesis, providing novel insights into the current prevention and treatment of this disease and related birth defects.

Protein-protein interaction (PPI) participates in almost all important biological processes in the organism and plays a vital role in the basic life process of cells. It is of great biological significance to develop a high-throughput PPI detection method. At present, the next-generation sequencing (NGS) technology has developed rapidly and can determine more than 1 billion template DNA sequences in a few days. Due to the unique sensitivity, specificity, high throughput, and multiplexing advantages of parallel DNA sequencing technology, it has been used as a broad-spectrum molecular counter and applied to genome sequencing, transcriptome sequencing and other fields. The nucleic acid barcode technology realizes the high-throughput detection of PPI by connecting the oligonucleotide tag with the target proteins to mark the encoded protein, and then using the high-throughput sequencing method to detect the interacting protein. This technology promotes the rapid development of PPI detection methods, improves the throughput of single experiment detection, and provides a strong technical support for building PPI network. In this paper, we describe in detail the design, generation and reading of nucleic acid barcodes in the PPI detection method. By analyzing the application examples of the nucleic acid barcode technology in PPI research, this paper discusses their advantages and disadvantages, evaluates the reliability of data, and discusses the future development trend of PPI detection methods based on the nucleic acid barcode technology.

Lymphocyte cytoplasmic protein 2 (LCP2) is an adaptor protein that participates in the T cell receptor signal transduction pathway through activating downstream signaling factors to complete the immune response. LCP2 also plays an important role in the occurrence, development and metastasis of malignant tumors. The high expression of LCP2 leads to adverse prognostic effects and reduces patient survival rates. The specific mechanisms of these reactions involve multiple signaling pathways. Here we introduce the structure and basic functions of LCP2, and review the important roles that LCP2 plays in the occurrence and development of malignant tumors through participating in NF-κB, MAPK, JAK/STAT, and PD-1/PD-L1 signaling pathways. The potential role of LCP2 as a tumor treatment target was also discussed for providing a theoretical basis and reference for the diagnosis, treatment, and biomarker screening of relevant diseases.

Induced pluripotent stem cell-derived mesenchymal stem cells (iMSC) have been proposed as an alternative source to primary mesenchymal stem cells(MSC), showing multiple advantages in disease treatment research. However, it remains unclear which disease type iMSCs are more suitable and whether they carry potential risks. This study utilized high-throughput sequencing data from public databases to compare iMSC with bone marrow-derived MSC (BM-MSC), adipose-derived MSC (AD-MSC), and umbilical cord-derived MSC (UC-MSC). Through various bioinformatics methods, including differential gene expression analysis, functional enrichment analysis, protein-protein interaction network analysis, and CMap database screening. The results indicated that iMSC possesses unique gene transcription characteristics, showing significant differences in gene expression compared to the three commonly used MSC types, particularly in genes related to neural and muscle development and immune regulation. Functional enrichment analysis of the differential genes further confirmed iMSC's potential advantages in treating neuro-related diseases, along with its lower immunogenicity but higher tumorigenic risk. Analysis using the CMap database identified potential gene targets and small molecule inhibitors to mitigate the tumorigenic risk of iMSC, providing possible strategies to reduce the risks associated with iMSC application. In summary, as a potential source for cell therapy, iMSC shows promising advantages in treating neurological diseases, but its safety needs to be validated through further experiments and clinical studies.

Congenital heart disease (CHD) refers to congenital structural abnormalities of the heart, including defects in the myocardial wall, valves, and major blood vessels. While genetic factors such as mutations and aberrant gene expression during embryogenesis contribute to CHD, they only account for part of cases. There are more and more studies on epigenetic histone modifications in CHD, suggesting that they are increasingly important in the pathogenesis of CHD. With the development of mass spectrometry-based proteomics technology, a spectrum of novel histone post-translational modifications, such as succinylation, glycosylation, lactylation, and β-hydroxybutyrylation, have been uncovered to contribute to various diseases. However, it remains unclear how these novel modifications regulate gene expression and pathological processes during the development and progression of CHD. This article will delve into the mechanisms of various histone modifications that regulate genes associated with cardiac development. It will approach the topic from both the perspective of classic histone modifications and novel histone modifications, aiming to unveil the significance of histone-driven epigenetic mechanisms in the etiology of CHD. Furthermore, it seeks to offer insights into the etiology of CHD, providing a theoretical basis for clinical treatment and timely prevention of this condition.

Circular RNA (circRNA) is a special class of non-coding RNA (ncRNA) in eukaryotic cells, which is highly stable. At present, circRNA has been widely found to be involved in the occurrence and development of various diseases as microRNA (miRNA) sponges. As a research hotspot in recent years, the function and effects of circRNA are constantly being discovered and utilized. CircRNAs can act as protein scaffolds to bind targets more tightly and regulate transcription of disease signaling molecules, as well as participate in protein expression as translation templates. A substantial body of research has established the pivotal role of ferroptosis in disease development. Iron and lipid metabolism-related targets are recognized as key components in the regulation of ferroptosis. Recent studies have revealed that circRNA not only participates in ferroptosis regulation as a miRNA and protein scaffold but also exerts control over ferroptosis-related signaling molecules at the transcriptional level. Currently, synthetic circRNA molecules have been engineered for the development of RNA vaccines. Additionally, a range of gene therapy technologies and drug delivery vectors offer a promising foundation for the development of therapeutics targeting circRNA. This paper describes the functions and effects of circRNA, its role as a key target in the regulation of ferroptosis, and its implications in various diseases. Special emphasis is placed on discussing the role of circRNA in multisystem diseases, including tumors, nervous system disorders, and diabetes-related complications. Additionally, drug development strategies related to circRNA are summarized to provide insights for the development of therapeutic drugs targeting ferroptosis.

Protein glycosylation, one of the most common post translational modifications (PTMs) of proteins, is widely present in living organisms. In eukaryotic cells, glycosylation modifications have a significant impact on protein folding, conformation, distribution, stability and activity. The glycosylation of glycoproteins is crucial for maintaining the order of interactions between all differentiated cells in multicellular organisms. The dysfunction of protein glycosylation may lead to the occurrence of diseases and development of infectious diseases. So far, changes in protein N/O-glycan have been identified as biomarkers for the development of tumors and certain infectious diseases. Therefore, this article reviews the glycosylation of major immune molecules such as B-cell receptor (BCR), T-cell receptor (TCR), cytokines (CK), complement and immunoglobulin (Ig), as well as the relationship between these glycosylation and infectious diseases. The aim is to understand the association between glycosylation of immune molecules and infectious diseases, and to provide new ideas and strategies for the treatment of infectious diseases.

Lysosomal storage diseases (LSD) are a category of genetic metabolic disorders, originating from genetic mutations in lysosomal acid hydrolases, leading to enzymatic deficiencies. Consequently, this triggers an abnormal accumulation of biological macromolecules within lysosomes, subsequently causes significant damage to cellular, tissue, and organ functions. Mutations or deficiencies in β-galactocerebrosidase (GALC) result in the accumulation of psychosine, causing progressive demyelination and triggering Krabbe Disease, a form of neurodegenerative lysosomal storage disease. The specific mechanisms involved in disease regulation are not yet fully elucidated. Increasing reports of pathogenic mutations in the GALC gene, coupled with analyses of GALC protein structure, have gradually enhanced the understanding of how GALC mutations contribute to Krabbe Disease. This knowledge offers robust evidence for the development of potential therapeutic drugs. Furthermore, GALC plays a dual role in various tumor processes, acting as a tumor suppressor in some cancers while acting as a carcinogen in others. However, a comprehensive analysis of GALC’s impact on cancer requires further in-depth research, offering insight into GALC as a potential target for tumor promotion or suppression. GALC is also associated with various neurodegenerative diseases, including Alzheimer’s disease, Parkinson’s disease, and multiple sclerosis. Due to the complexity of GALC’s mechanisms, current treatments for Krabbe Disease caused by GALC deficiency primarily involve single-modality and multi-modality therapies. Nevertheless, developing truly effective treatments necessitates deeper research into the pathogenic mechanisms arising from GALC gene defects. This review summarizes the structural and functional characteristics of GALC, and discusses its roles in the development of the nervous system and tumorigenesis, as well as the latest advances in related research. The aim is to lay the theoretical foundation and provide references for exploring GALC’s regulatory mechanisms and developing innovative drugs for treating associated diseases in the future.

Fostering virtue through education is the fundamental task of higher education, while the performance of ideological and political education is critical to achieving this task. “Biological Basis”, the core course of the bioengineering major which has a close relationship with other required courses, plays an important role in the professional training system. From the aspects of the necessity of ideological and political construction of “Biological Basis”, the exploration and design of ideological and political elements, the practice of ideological and political teaching, the optimization of ideological and political evaluation systems, and the effect of ideological and political teaching, this study elaborated on the constructing process of “Biological Basis”. The ideological and political elements were explored from celebrity anecdotes, daily life, research cases, and current events were further integrated into the teaching process of “Biological Basis” by the establishment of a teaching team and the online and offline mixed teaching methods, the development of case teaching, and the implementation of group discussion. Moreover, the ideological and political course teaching-effect evolution system was established. The results showed that students' interest in learning and the average score of the final paper significantly improved, from 66 points or less before the curriculum reform to 76 points after the reform. In addition, more than 92% of the students thought that the ideological and political teaching of “Biology Basis” was good, with a course satisfaction of 98%. The course's ideological and political teaching improves the quality of “Biology Basis”, achieving the goal of cultivating students' professional knowledge and comprehensive quality together. The construction experience of this course will provide some references for the ideological and political teaching and reform of other biology courses.

Type 1 diabetes is caused by impaired function of pancreatic β-cells and insufficient insulin secretion. Currently, it is primarily managed with exogenous insulin supplementation; however, exogenous insulin cannot precisely regulate blood glucose levels, and severe hypoglycemia can be life-threatening. Islet transplantation serves as an alternative therapy, but faces challenges such as a shortage of organ donors and the risk of cross-species infections from xenogeneic sources of islet β-cells. Thus, obtaining a sufficient and safe supply of islet β-cells remains a significant challenge in cell therapy for type 1 diabetes. This study aims to differentiate human induced pluripotent stem cells (hiPSCs) into islet β-cells in vitro, offering a potential new strategy for treating type 1 diabetes. To achieve this, we utilized a differentiation strategy that combines 2D and 3D culture systems to simulate the in vivo developmental environment of islet β-cells and employed various growth factors to regulate key signaling pathways crucial in pancreatic development and β-cell differentiation, including the Notch, Wnt, and TGF-β/Smad signaling pathways. Our results show that under combined 2D and 3D culture conditions, the expression of specific genes at the stages of definitive endoderm, pancreatic progenitors, pancreatic endocrine cells, and islet β-cells significantly increased (P<0.05). And there was a marked enhancement in insulin content and secretion following glucose stimulation (P<0.05). In summary, this study successfully established a differentiation strategy from hiPSCs to functional islet β-cells, providing a new cell therapy approach for type 1 diabetes. This method not only offers new tools for studying the developmental biology of islet β-cells, but also provides a potential source of islet β-cells for clinical applications, potentially overcoming the limitations of current treatment methods.

With the acceleration of population aging, all kinds of cardiovascular diseases caused bycardiac aging have become a health problem that cannot be ignored. In the heart, about 95% of ATP come from the cardiomyocytes to maintain the pumping function. Mitochondrial dysfunction can lead to myocardial energy deficiency, cardiomyocytes damage and death, or myocardial senescence. Therefore, the intact function of mitochondria plays an important role in maintaining the normal function of the heart and is considered as a key feature of cardiac aging. This paper reviews cardiac aging and mitochondrial dysfunction, and mainly summarizes the characteristics of aging heart and the changes in mitochondrial structure and function of senescent cardiomyocytes. We focus on the five major factors leading to cardiac aging caused by mitochondrial dysfunction, including changes in the mitochondrial numbers and morphology, mitochondrial DNA mutations, mitochondrial quality control failures, mitochondrial enzyme changes, and mitochondria-related metabolites and stress signals changes. We also summarize the treatment methods and mechanism of cardiac aging by targeting mitochondria, anddiscuss the current status and future direction of mitochondrial therapy for cardiac aging.

Pseudouridine (Ψ)-modified in vitro-transcribed mRNA (Ψ-IVT-mRNA) is used in two licensed SARS-CoV-2 mRNA vaccines in response to the COVID-19 pandemic worldwide. Recently the Ψ-IVT-mRNA has been demonstrated to cause ribosome stalling and lead to +1 ribosomal frameshifting both in vitro and in vivo, resulting in aberrant protein products. The aberrant proteins may have the potential to induce off-target T-cell or antibody responses and other unintended side effects in people vaccinated with SARS-CoV-2 mRNA vaccines. The finding highlights a key aspect of developing mRNA-based vaccines/therapeutics with improved outcomes.

Vasculogenic mimicry (VM) refers the process by which tumor cells form new microcirculation patterns in the absence of endothelial cells. Clinically, VM is associated with an aggressive phenotype and poorer patient survival. Many previous studies have focused on angiogenic factors. Recently, more and more studies have found that the extracellular matrix around tumor cells plays a significant role during tumorigensis. Extracellular matrix proteins may render tumor cells to form vasculogenic mimicry. In this study, we investigated the role of laminin (LN) matrix in tumor progression and VM formation in melanoma and osteosarcoma. Compared with the control group, the adhesion, proliferation, migration, stemness expression and VM formation ability of cancer cells were significantly enhanced under high concentrations of LN substrate culture. Immunofluorescence staining and real-time quantitative PCR were used to analyze the internal molecular mechanism. The results showed that compared with the control group, the laminin matrix significantly promoted the expression of CD133 in tumor cells, and the mRNA expression of E-cadherin is down-regulated(P<0.05), while the mRNA expression of Integrin, N-cadherin, Vimentin, Snail and Twist is up-regulated(P<0.05). These results indicate that laminin promotes the adhesion, proliferation, migration and VM formation of tumor cells, and reveals the potential mechanism of VM formation, providing a new idea for the VM diagnosis and treatment.

Cellular senescence refers to the stable state of cell cycle arrest in which cells lose the ability of division and proliferation. Various intracellular and extracellular stimuli can induce cell senescence. Senescent cells exhibit multiple hallmarks, such as the upregulation of cell cycle inhibitor proteins p16^INK4a and p21^Cipl, DNA damage responses, and structural and metabolic alterations. Another major hallmark of senescent cells is that they express and secret a variety of factors, including cytokines, chemokines, growth factors, proteases, and other bioactive molecules, defined as the senescence-associated secretory phenotype (SASP). SASP factors exert multiple biological functions through the cell autonomous autocrine manner or cell non-autonomous paracrine fashion. In this review, we summarize the composition of SASP, and point out its high heterogeneity and dynamics. We then summarize the regulatory mechanisms of SASP at various levels including transcription, post-transcription, translation, post-translational modifications, and epigenetics. Afterwards, we summarize the various biological functions of SASP, including its beneficial effects in tumor suppression, tissue repair, and embryonic development, as well as its detrimental effects in inducing cell senescence, promoting tumor occurrence and development, age-related diseases, and organismal aging. We further discuss the potential applications of the SASP, which overview the senolytic therapy for selectively clearing senescent cells and senomorphic therapy for inhibiting SASP to intervene age and age-related diseases. Finally, we outline several challenges in identifying and detecting senescent cells and SASP factors in vivo, and provide some practical recommendations and new techniques to address these challenges.

Breast cancer is one of the malignant tumors that seriously affects women’s lives and health. In recent years, the anti-tumor effect of artemisinin (ART), an active ingredient of traditional Chinese medicine, has become more prominent. However, the effectiveness of ART in the treatment of breast cancer metastasis and its mechanism are still unclear. In this paper, we used the mouse breast cancer cell 4T1 and mouse mammary epithelial cell HC11 to explore the role of ART in inhibiting metastasis and inducing apoptosis. The MTS assay showed that ART was able to significantly inhibit the viability of 4T1 cells in a concentration andtime-dependent manner. Analysis of cell colony formation, Transwell and cell scratch results revealed that ART significantly inhibited the metastasis of breast cancer 4T1 cells. Flow cytometry results showed that ART promoted ROS production, loss of mitochondrial membrane potential, cell cycle arrest and thus apoptosis in tumor cells. Bioinformatic analysis methods and studies confirmed that extremely significant high expression of TIGIT (P < 0.001) was closely associated with breast cancer metastasis, and that ART was able to significantly reduce the gene expression level of the TIGIT/CD155 signaling axis (P < 0.001, P < 0.01). In summary, this study reveals the regulatory mechanism by which ART may inhibit breast cancer metastasis by blocking the TIGIT/CD155 signaling axis, which provides a basis for ART as a potential therapeutic agent for breast cancer, and contributes to the exploration of a new way of treating tumors using the active ingredients of Chinese herbal medicine.

DNA damage triggers cells to initiate a series of DNA damage response (DDR), including DNA damage repair, cell cycle checkpoint activation, cell cycle arrest, activation of various intracellular signal transduction pathways, and cell apoptosis, etc. DNA Damage Repair, an important mechanism by which cells maintain genomic stability, was awarded the Nobel Prize in Chemistry in 2015. DNA damage repair pathways mainly include: base-excision repair (BER), nucleotide excision repair (NER), mismatch repair (MMR), homologous recombination (HR) and non-homologous end joining (NHEJ). They play an important role in the repair of DNA damage such as single-strand break (SSB) or double-strand break (DSB). DNA damage repair defects are closely related to tumorigenesis and development and is also an important target for tumor therapy. Poly-ADP-ribose polymerase (PARP) and breast cancer susceptibility gene BRCA1/2 and others in the DNA damage repair pathways have synthetic lethality effects. It makes PARP inhibitor (PARPi) be the first and currently the only commercially available synthetic lethal target drug for tumor therapy. PARPi has good efficacy in the treatment of ovarian cancer and a variety of solid tumors, which make the research and development of synthetic lethal target drugs related to DNA damage repair and DDR pathway become a hot spot. Other targets under research mainly include: ataxia telangiectasia-mutated protein (ATM), ataxia telangiectasia and RAD3 related protein (ATR), DNA-dependent protein kinase catalytic subunit (DNA-PKcs), checkpoint kinase1 (CHK1), Checkpoint kinase 2 (CHK2), mitogen-preventing protein kinase WEE1,etc. The combination of PARPi with other DDR target drugs, anti-angiogenesis drugs or immune checkpoint inhibitors may become effective means and development prospect to overcome PARPi resistance and improve the therapeutic effect. Here we review the key molecules and potential tumor therapeutic targets in DNA damage repair and related DDR pathways, and the research on synthetic lethal targets and their application and prospect in ovarian cancer. We aim to provide guidance for basic research and clinical application.

p53 is an important tumor suppressor in the body, but research has found that p53 is one of the most frequent alterations in nearly half of human cancers. Mutant p53 (mtp53) not only promotes tumor development but also serves as a pivotal factor in tumor immunity. Research has demonstrated that mtp53 participates in many aspects of tumor immune response through multiple mechanisms. We firstly describe the structure and function of mtp53, then the molecular mechanisms of mtp53 in tumor immunity, including four aspects of promoting tumor immune escape, facilitating oxidative stress, regulating tumor-associated inflammatory signaling networks and tumor-infiltrating immune cell recruitment and differentiation. Finally, we summarize the effects of mtp53 on the immune micro-environment of lung cancer, triple-negative breast cancer and colorectal cancer as well as the advances in the application of mtp53 in the treatment of cancers with PD-1/PD-L1 inhibitors, which may provide suitable directions and options for the immunotherapy of mtp53-expressing cancers.