At the nanoscale, metals exhibit special electrochemical and optical properties, which play an important role in nanobiosensing. In particular, surface plasmon resonance (SPR) based on precious metal nanoparticles, as a kind of tag-free biosensor technology, has brought high sensitivity, high reliability, and convenient operation to sensor detection. By applying an electrochemical excitation signal to the nanoplasma device, modulating its surface electron density, and realizing electrochemical coupling SPR, it can effectively complete the joint transmission of electrical and optical signals, increase the resonance shift of the spectrum, and further improve the sensitivity of the designed biosensor. In addition, smartphones are playing an increasingly important role in portable mobile sensor detection systems. These systems typically connect sensing devices to smartphones to perceive different types of information, from optical signals to electrochemical signals, providing ideas for the portability and low-cost design of these sensing systems. Among them, electrochemiluminescence (ECL), as a special electrochemically coupled optical technology, has good application prospects in mobile sensing detection due to its strong anti-interference ability, which is not affected by background light. In this review, the SPR is introduced using nanoparticles, and its response process is analyzed theoretically. Then, the mechanism and sensing application of electrochemistry coupled with SPR and ECL are emphatically introduced. Finally, it extends to the relevant research on electrochemically coupled optical sensing on mobile detection platforms. Metal nanoparticles were first applied to the Lycurgus cup in Rome in the 4th century AD. By embedding gold (Au) and silver (Ag) nanoparticles in glass, they can absorb and scatter specific regions of the visible light spectrum, presenting bright colors . Due to The interaction between light and noble metal nanoparticles generates a collective oscillation of conduction band electrons, which is called SPR. Only materials with a negative real dielectric constant and a positive virtual dielectric constant can better support surface plasmon polaritons, and the most commonly used precious metal nanoparticles are Au and Ag nanoparticles ,3. When R represents the radius of the particle, mε is the dielectric constant of the surrounding medium, and N represents the electron density, λ represents the wavelength of incident light, r + iεiε = ε refers to the complex dielectric constant of a metal, χ determined by particle shape. Therefore, factors such as the shape of nanoparticles, the wavelength of incident light, the type of material, and the surrounding medium can all affect the absorption and scattering processes.In order to understand the factors affecting the increase in absorption and scattering during plasma resonance, the Mie theory was proposed. Mie theory is the analytical expression of Maxwell equations with spherical boundary conditions that is used to describe the extinction spectrum of a given nanoparticle. In order to more accurately calculate the dielectric constant at different wavelength values and extend this theory to more complex nanoparticle shapes, a modified wavelength form of Mie theory was proposed ,10,11. Am is the refractive index sensitivity, Δn is the refractive index change caused by the adsorbent. d refers to the thickness of the effective adsorption layer, and dl refers to the attenuation length of the electromagnetic field.In the field of biosensors, when biological analytes are combined with the surface of nanoparticles, the refractive index of the surface of the nanoparticles will change, thus changing the movement of the SPR peak position. Among them, Ag nanoparticles have the highest negative dielectric constant among all plasma materials and are the most sensitive to changes in local refractive index. In addition, asymmetrically shaped nanoparticles are more sensitive to changes in the surface binding of biomolecules than spherical plasmas. The relationship between the position of the SPR peak and the binding of the adsorbent is described in Formula (2), where n) and the degree of surface binding between the analytes and the nanoparticles (d). Due to the sensitivity of the refractive index being the key to detecting target biomolecules, many studies in recent years have attempted to make the nanoparticle matrix highly sensitive to changes in refractive index. The relevant simulation results show that when the SPR peak of the material moves from the blue wavelength to the red wavelength, the sensitivity of the refractive index increases in a linear way . T. T89]. TAlthough this article has conducted relevant research on nanobiosensing motion detection based on electrochemically coupled optical technology, there are still some problems that need to be solved to further promote the application of electrochemically coupled optical technology in biosensing motion detection. First, the strictness of anti-interference requirements limited to optical sensing has greatly increased the complexity of optical path construction, limiting the realization of electrochemical coupling SPR resonance on smartphones. In addition, currently, nanoarray devices only achieve single biological modifications, only utilizing the improvement in sensing sensitivity brought by their nanostructures, and do not fully utilize the advantages of their array structures. Further exploration is needed to achieve multi-channel detection through the modification of different sensitive layers, enriching the output signals of nanoarray sensors. Secondly, this review has demonstrated the excellent electrochemical/optical sensing properties of nanomaterials. Then the biosensor characteristics of metal carbide nanomaterials with a two-dimensional layered structure, such as graphite oxide derivatives and MXene, need to be further studied to broaden the types of detection probes and promote the development of biosensors. However, the limited sensing characteristics brought by nanomaterials cannot fully meet the demand for biosensing specific detection. Therefore, further research is needed to improve the sensing selectivity of nanomaterials by coupling them with biologically sensitive layers such as proteins, peptides, or DNA. Finally, this study conducted a series of detailed studies on ECL detection based on smartphones. The sensing system is sufficiently complete to meet the relevant biological sensing and detection needs; however, there are still many aspects that can be improved with the development of ECL technology. Although the luminescent material is not consumed in the reaction, it is difficult to recycle and reuse. Solid state luminescence not only facilitates detection operations but also reduces testing costs by fixing and modifying the luminescent material on the electrode surface, which is conducive to mobile detection applications. Although current research utilizes mobile phone cameras to dynamically capture luminous videos, the luminous processing is still based on luminous imaging and does not directly analyze the optical signal. Therefore, real-time processing of luminous signals using smartphone light sensors and other components is another area that needs improvement. In the future, there is still a long way to go for related in-depth research, such as integrating more precise sensors on smartphones or developing more precise detection technologies to achieve quantitative analysis of fingerprint substances and promote mobile biosensing detection during the unlocking process of mobile phones. Given the excellent performance of ECL in imaging analysis, further exploration can be made in the spatial distribution of luminescence, such as multicolor luminescence. At the same time, the convenience of image processing using a smartphone can enable multi-channel biosensing detection applications on multicolor luminescent materials.(1)Research on multi-channel biosensing detection based on nanoarray sensors. Precious metal nanoarray sensors were prepared to realize electrochemical coupling SPR detection and modify multiple biosensitive layers on them. Build a multi-channel electrochemically coupled optical detection platform based on nanoarray sensors. Realize simultaneous detection of multiple target analytes.(2)Research on sensing detection based on a nanomaterial composite biosensitive layer studies the electrochemical/optical sensing characteristics of metal carbide materials with a two-dimensional layered structure. Preparation of composite probes for nanomaterials and biosensitive layers to achieve specific biosensing detection of target analytes.(3)Research on biosensing detection based on quantum dots in solid state ECL built a dynamic real-time ECL analysis system based on smartphone light sensors. And on this system, study the ECL-related characteristics of new quantum dot nanomaterials and complete the construction of solid-state ECL sensing components. Implement relevant biosensing detection applications.(4)Research on multicolor luminescent biosensing detection based on smartphone preparation of potential resolved multicolor ECL-sensitive probes and construction of multi-channel biosensing components. Build an ECL platform based on smartphones and conduct multimodal analysis of luminescence signals. Furthermore, mobile sensing for the detection of complex biological samples can be achieved.The ideas for future work are:This paper mainly introduces the basic concepts of local SPR, electrochemically coupled local SPR resonance, ECL, and related biosensor detection applications. Firstly, the local SPR is induced by nanoparticles, and its sensing mechanism is briefly introduced. Then the role of electrochemistry in improving local SPR signals is introduced, and the mechanism of electrochemical coupling of local SPR and its applications in biosensors are emphatically introduced. In order to meet the growing demand for mobile detection, ECL technology is introduced, and its luminescence mechanism, luminescent materials, and applications are described in detail. Finally, from the perspective of real-time detection applications, a detailed analysis was conducted on the miniaturization and intelligent development of ECL excitation and luminescent detectors.


Cancer is a huge challenge for people worldwide. High reactive oxygen species (ROS) levels are a recognized hallmark of cancer and an important aspect of cancer treatment research. Abnormally elevated ROS levels are often attributable to alterations in cellular metabolic activities and increased oxidative stress, which affects both the development and maintenance of cancer. Moderately high levels of ROS are beneficial to maintain tumor cell genesis and development, while toxic levels of ROS have been shown to be an important force in destroying cancer cells. ROS has become an important anticancer target based on the proapoptotic effect of toxic levels of ROS. Therefore, this review summarizes the role of increased ROS in DNA damage and the apoptosis of cancer cells caused by changes in cancer cell metabolism, as well as various anticancer therapies targeting ROS generation, in order to provide references for cancer therapies based on ROS generation. In the 21st century, cancer has become one of the most important diseases threatening human life and health. Lung cancer is the most common cancer type and the leading cause of death in both male and female patients. In contrast, breast cancer is most common in women, accounting for 11.7% of all female cancer cases in 2020 ,4,5. GloCompared with normal cells, cancer cells show remarkable features, such as continuous proliferation, abnormal DNA damage response (DDR), and high levels of ROS . The occCompared with normal cells, cancer has the characteristics of cellular metabolic reprogramming, that is, cancer cells adapt metabolism to meet their own nutritional and material needs for continuous proliferation, thus promoting transformation and tumor development . In receUsnea barbata extract in canola oil showed anticancer potential through ROS generation leading to DNA damage and autophagy [All cancer cells have obviously high levels of ROS, but excessive accumulation of ROS can aggravate oxidative damage of the DNA of cells and induce the apoptosis of cancer cells. Under normal circumstances, an appropriate amount of ROS is one of the necessary conditions for maintaining cell signaling and redox homeostasis, but a further increase in ROS levels due to exogenous stimulation or metabolic changes in cancer cells is carcinogenic. High levels of ROS in cancer cells put cancer cells in a state of high oxidative stress and lead to the oxidative stress-induced damage of proteins, lipids, DNA, and mitochondria , among wutophagy . TherefoOver the past few decades, the relationship between oxidative stress, ROS, and cancer has been elucidated. Current research reviews on cancer mainly focus on the adaptive metabolism of cancer cells and cancer development, the abnormal metabolism of cancer cells and the shaping of the tumor immune microenvironment, and the relationship between metabolic phenotypes and cancer. However, the relationship between ROS accumulation caused by cancer metabolism and DNA damage and the apoptosis of cancer cells has not been systematically elaborated. Therefore, this review article aims to systematically summarize the dual mechanism of promoting and inhibiting cancer via ROS accumulation in cancer cells and clarify how ROS levels induce DNA damage and promote the apoptosis of cancer cells. In addition, the various cancer treatment strategies targeting ROS generation that have been used in recent years are also summarized to provide references for further research on drugs, specifically with respect to targeting ROS generation in cancer cells or anticancer therapies.Both exogenous environmental stimuli and endogenous factors can lead to ROS generation. When exposed to exogenous stimuli such as air pollutants, heavy metals, radiation, and anticancer drugs, ROS will abnormally accumulate in cells, leading to oxidative stress and gene mutations, which may induce the transformation of normal cells into cancer cells . Air pol2−), hydroxyl radical (−OH), hydrogen peroxide (H2O2), and lipid hydroperoxide (LOOH). ROS mainly comes from mitochondria [2− and maintains the proliferative phenotype of colon cancer cells [2O2 leads to extracellular oxidative stress and promotes tumor growth and invasion [The metabolic reprogramming characteristic enables cancer cells to generate ROS at higher levels than normal cells through a variety of biological pathways. In tumor tissues, the rapid energy metabolism and proliferation of tumor cells generate a large number of ROS, including superoxide  activation and shifts metabolism from glycolysis to oxidative phosphorylation [The abnormal metabolic behavior of cancer cells is to support the need for cancer cells to maintain their development in harsh environments such as lack of oxygen and nutrition . Becauserylation . OverallAn imbalance between redox homeostasis and oxidative stress will inevitably result from the accumulation of abundant metabolites from the hyperactive metabolic activities of cancer cells. Continuous oxidative stress will lead to a large accumulation of ROS within cancer cells, and the effect of ROS on cancer progression has been shown to be paradoxical ,80,81. HThe moderate accumulation of ROS is conducive to tumor maintenance and development . Increas2− acts as a positive regulator of the Rac1-PI3K signaling pathway, which is associated with stimulating the migration and invasion of Nex10C mouse melanoma cells [Many research studies have shown that excessive ROS generation has a pro-cancer effect ,87. Highma cells . On the ma cells . Furtherma cells . The aboSince ROS levels play a role in promoting cancer, a further increase in ROS levels can promote antitumor signaling and trigger oxidative stress, thus inducing tumor cell apoptosis through the pro-apoptotic pathway. The imbalance of ROS induces adverse cancer outcomes by affecting signal transduction within the DDR  because Apoptosis is an important homeostatic mechanism of the body that plays an important role in maintaining the homeostasis of normal cells. Cancerous cells can break through the age limit and have the ability to continue to proliferate, which is also the reason for the occurrence of tumors. The apoptosis of cancer cells allows for the development of tumors to be controlled, which also brings greater hope to cancer treatment. At present, there are three major apoptotic pathways: (i) the endogenous mitochondrial apoptosis signaling pathway, (ii) the death receptor pathway, and (iii) the endoplasmic reticulum signaling pathway. It has been demonstrated that ROS generated in cancer cells induces apoptosis through the activation of these apoptotic signaling pathways and plays a role in tumor inhibition ,99.Toxic levels of ROS can activate members of the mitogen-activated protein kinase (MAPK) family, among which the activation of the c-Jun N-terminal kinase (JNK) signaling pathway  can lead to intrinsic or extrinsic apoptotic signaling to initiate apoptosis ,100. ResAmong the many apoptosis signaling pathways, the mitochondrial apoptosis signaling pathway plays an irreplaceable role in cancer cell apoptosis. Mitochondria are not only the “energy factories” of cells, but also the regulatory centers of cell survival and death. Mitochondrial dysfunction leads to excessive ROS generation, which leads to oxidative stress and aggravates mitochondrial dysfunction, leading to apoptosis. In addition, when cytokines, death receptors, cellular metabolites, and other factors stimulate mitochondria and affect the mitochondrial electron transport chain and energy metabolism, it is likely to activate the endogenous mitochondrial apoptosis signaling pathway, leading to cell apoptosis. Caspases play a special role in the mitochondrial intrinsic apoptosis signaling pathway and death receptor apoptosis signaling pathway, and its activation is one of the signs of cell apoptosis. ROS causes mitochondrial membrane damage and releases cytochrome c into the cytoplasm, which forms a complex with apoptotic protein activating factor 1 (Apaf-1) and pro-caspase-9 to induce the cleavage of caspase-3 and caspase-7, eventually leading to DNA fragmentation and cell death ,104. In It has been confirmed that ROS-induced apoptosis is related to endoplasmic reticulum stress ,108. TheCancer cells employ multiple adaptive mechanisms in response to elevated ROS and oxidative stress . In ordeAs we all know, endogenous ROS mainly comes from mitochondria and is a by-product of the oxidative phosphorylation of mitochondria. Research studies have demonstrated that uncontrolled mitochondrial ROS (mROS) affects normal cell survival and carcinogenic transformation . HoweverThe maintenance of redox homeostasis in cancer cells under high ROS levels supports cancer cell survival and proliferation. Cancer cells can make adaptive changes to oxidative stress, such as metabolic reprogramming, enhanced antioxidant capacity, and resistance to apoptosis, to meet the needs of their own continuous growth. As tumor-initiating cells, cancer stem-like cells (CSCs) have a more powerful antioxidant defense system to counteract, clear, and maintain tumorigenesis . ResearcThe antioxidant response is one of the key cellular defense mechanisms used by cancer cells to prevent oxidative stress, and nuclear factor erythroid 2-related factor 2 (Nrf2) is the core of the antioxidant reaction . Nrf2 acCancer cells adapt to hypoxia through changes in energy metabolism. The maintenance of the continuous high proliferation state of cancer cells requires the consumption of a large amount of oxygen, so the hypoxic environment is considered to be a sign of the tumor microenvironment . HypoxicThe occurrence and progression of cancer are closely related to bidirectional regulatory mechanisms below or above the threshold ROS level. Cancer cells also have the sufficient adaptive capacity to cope with the high level of ROS pressure and can maintain their survival and development through a variety of adaptive mechanisms, which is a favorable obstacle to tumor treatment.Recently, cancer treatment strategies that target ROS regulation in cancer cells have attracted increasing attention. The occurrence of cancer is a serious threat to human health. The current cancer treatment methods include radiotherapy, chemotherapy, surgery, immunotherapy, and molecular targeted therapy, and their cancer treatment effects have also been recognized in the clinical treatment of cancer patients. However, due to non-specific targeting, various adverse reactions, and the continuous emergence of tumor drug resistance, the application of traditional anticancer strategies has been greatly limited . High leCurrently, ROS-generation-based cancer treatment strategies have been widely used. Anticancer drugs are the traditional methods of cancer treatment. Some anticancer drugs are based on the inhibition of cancer cell proliferation and cell death caused by ROS generation, which has advantages over the resistance-based shortcomings of traditional drugs (such as 5-fluorouracil and cisplatin) . On the In addition to pharmacotherapy, ROS generation is also widely used in several novel anticancer therapies, such as photodynamic therapy (PDT), sonodynamic therapy (SDT), chemodynamic therapy (CDT), and the combination of multiple therapeutic therapies.PDT is a new type of anticancer therapy in which ROS generated during treatment induces cell death through oxidative damage to cells . Existing research studies show that PDT has a good effect on the treatment of cervical cancer, colon cancer, skin cancer, and lung cancer and that it has fewer adverse effects than surgery, chemotherapy, and radiation treatment ,142. PDT2O2 [Although PDT has the ability to accurately target ROS generation, it has certain limitations, such as limited light penetration depth, which is not ideal for the treatment of deep tumors . SDT use2O2 . OverallPro-oxidative anticancer agents (PAAs) targeting redox imbalance are effective for the treatment of cancer patients . PAA is However, compared with contemporary conventional anticancer therapies and some other novel anticancer therapies, anticancer therapies targeting ROS generation within cancer cells have unique advantages but also certain limitations. Traditional surgery and chemotherapy drugs may have good efficacy in the early treatment of cancer, but the effectiveness of the treatment of malignant tumors is insufficient, and there are side effects and a high risk of recurrence. Cancer cells are resistant to treatments such as chemotherapy and radiotherapy and cannot be eradicated because of their properties such as rapid metabolic adaptation, high proliferation, and other resistance mechanisms. Cancer stem cells (CSC) exert resistance in radiotherapy and chemotherapy, which is the resistance mechanism of cancer treatment. For example, breast CSCs have traditional hyperthermia and radiation resistance . PhotothAdaptive metabolic alterations lead to the abnormal accumulation of ROS in cancer cells, and cancer cells rely heavily on ROS levels to enhance tumorigenesis or induce apoptosis ,159. ThiIn clinical practice, in order to cope with the highly invasive and metastatic nature of malignant tumors, it is often necessary to take chemotherapy drugs for a long time. However, the long-term and excessive use of chemotherapy drugs will not only increase the risk of harmful side effects and drug resistance of cancer cells, but also cause disrupt the tumor immune microenvironment and immune escape. Therefore, traditional cancer drugs do not have an advantage in the ongoing battle against cancer. At present, PDT, SDT, and CDT, which are commonly used in clinical practice, have certain advantages in traditional cancer treatment. However, the treatment of highly drug-resistant tumors is still a major problem, often requiring the combination of two or more treatments. Cancer drug resistance is a headache, which means that many chemotherapy drugs are not effective enough to kill cancer cells, and anticancer drugs targeting ROS generation can cause the apoptosis of drug-resistant cancer cells. Patchouli alcohol has been shown to have potential in the treatment of non-small cell lung cancer with or without vincristine resistance by inducing ROS-mediated DNA damage and promoting cell apoptosis . HoweverA variety of anticancer therapies can induce the generation of ROS to trigger the apoptosis of cancer cells; however, at the same time, the intrinsic detoxification mechanism of the body against ROS-mediated oxidative stress will also be activated. Autophagy can remove oxidative damage of organelles and biomolecules and induce the drug resistance of cancer cells, thus creating resistance to the cytotoxic effect of ROS . The aggThe rapid metabolic adaptation of cancer cells occurs to meet the needs of endogenous growth, proliferation, and metastasis. The crosstalk between the complex pathways activated in cancer metabolic reprogramming has still not been fully elucidated, and the specific mechanism of metabolic changes regarding the generation and influence of high levels of ROS needs to be further elucidated. In addition, the specific mechanism of action of different ROS types in cancer cells in each stage of cancer development needs to be studied further, and the question of how to accurately characterize and locate each ROS type is also a problem to be solved. Traditional quantitative ROS detection techniques, such as chemical and immunological techniques, cannot meet the requirements of clinical detection standards. Therefore, the development of temporal- and spatial-specific ROS detection techniques to elucidate the complex biological mechanism of ROS-mediated redox in cancer is of great significance . The solDue to the inherent heterogeneity of tumors, current drug treatments and therapies are unable to meet the increasingly severe changes with respect to cancer deterioration situations, and with the continuous adaptation of malignant tumors to the hypoxic microenvironment and the development of drug resistance through changes in material and energy metabolism, other obstacles have become increasingly prominent, posing great challenges to clinical cancer therapy. ROS are effective anticancer targets, and anticancer therapies targeting ROS are proven to be effective means of cancer treatment. However, among the various therapeutic methods used to induce cancer cell apoptosis through ROS generation, the effectiveness and specificity of such treatments still need to be improved. Therefore, it is necessary to ensure that the generation of ROS with therapeutic effects is selective and specific for cancer-type cells. This should be the goal when developing anticancer drugs that target ROS generation. Therefore, based on the important role of ROS levels in the regulation of cancer, the wide application of anticancer therapy methods targeting ROS generation is promising for cancer treatment.


The use of performance-enhancing substances in bodybuilding is a well-known and long-standing issue. This comprehensive review article provides a detailed overview of the history of performance-enhancing substance use in bodybuilding, the types of substances commonly used, and the short-term and long-term health effects associated with their use. Additionally, the article discusses the regulation of performance-enhancing substances in various countries and the role of healthcare professionals in preventing substance use. The article also highlights the impact of substance use on society and the importance of prevention and intervention strategies. Finally, the article emphasizes the role of policymakers in addressing performance-enhancing substance use, including the development of regulations, penalties for violating rules, and the provision of resources for prevention and intervention programs. Overall, this review article sheds light on the dark side of bodybuilding and provides insight into the public health implications of performance-enhancing substance use. Bodybuilding is a sport that involves the rigorous training and development of the body's muscles through a combination of weightlifting, cardio, and nutrition. While it has been around for centuries, it gained popularity as a competitive sport in the 20th century, with the first Mr. Olympia contest in 1965. Since then, bodybuilding has become a global phenomenon with millions of enthusiasts and a billion-dollar industry ,4.Bodybuilding has a long history of performance-enhancing substance use. The early forms of PES in bodybuilding were simple remedies such as caffeine, alcohol, and opiates. However, the emergence of anabolic-androgenic steroids in the mid-20th century revolutionized the sport and transformed it into what we see today . In the Since then, the use of performance-enhancing substances in bodybuilding has evolved dramatically. Athletes now use various substances, including human growth hormone, insulin, diuretics, stimulants, and others, to gain a competitive edge. Additionally, the methods for administering these substances have become more sophisticated, including intravenous injections, transdermal patches, and oral dosages . The useThe objective of this review is to critically examine the use of performance-enhancing substances in the field of bodybuilding and shed light on the resulting implications for public health. By analyzing the prevalence, effects, motivations, and risks associated with using these substances, this article aims to understand the complex relationship between bodybuilding and performance-enhancing substances thoroughly. Through a comprehensive synthesis of current research and literature, the objective is to contribute to the existing knowledge base and promote informed discussions among professionals, policymakers, and the general public regarding the potential health consequences and broader societal implications of performance-enhancing substance use in the context of bodybuilding.Types of performance-enhancing substances used in bodybuildingAnabolic SteroidsAnabolic steroids are a class of synthetic substances that mimic the effects of the male hormone testosterone in the body. Bodybuilders and other athletes use these substances to increase muscle mass, strength, and endurance as well as to shorten recovery times after workouts. Anabolic steroids are also used medically to treat delayed puberty, muscle wasting, and osteoporosis . AnaboliLong-term use of anabolic steroids can lead to even more serious health consequences. It can cause kidney damage and increase the risk of prostate cancer. It can also lead to infertility, impotence, and reduced testicular size. Anabolic steroid use can also cause psychological effects, including mood swings, aggression, and depression . In addiHuman Growth HormoneHuman growth hormone (HGH) is a hormone naturally produced by the pituitary gland and plays a crucial role in stimulating growth, cell reproduction, and regeneration in humans. It is important in childhood for normal growth and development, and in adulthood, it helps to maintain muscle mass and bone density and regulate metabolism. HGH also significantly affects the immune system, cognitive function, and mood . HGH is Furthermore, studies have shown that long-term HGH use may increase cancer risk, particularly in the digestive system, such as colon, pancreas, and stomach cancer. This risk is likely due to the growth-promoting effects of HGH, which may stimulate the growth of cancerous cells ,6,15. ItInsulinInsulin is a hormone that plays a crucial role in regulating blood sugar levels. The pancreas produces it, and its primary function is to help cells in the body absorb glucose from the bloodstream, which is then used for energy. In bodybuilding, insulin is often used as a performance-enhancing substance due to its anabolic effects. Insulin stimulates the uptake of amino acids and glucose into muscle cells, promoting muscle growth and repair . HoweverAnother potential danger of insulin misuse is the development of insulin resistance, a condition where cells become less responsive to the effects of insulin. This can lead to elevated blood sugar levels and the development of type 2 diabetes . In addiDiureticsDiuretics are drugs that increase the rate of urine production by the kidneys, resulting in a loss of water and electrolytes from the body. In bodybuilding, diuretics are sometimes used to help reduce water retention in the body and promote a more defined and muscular appearance on stage. The use of diuretics in this manner is sometimes referred to as "cutting water weight" . HoweverMoreover, the use of diuretics can also put a significant strain on the kidneys. The kidneys are responsible for filtering the blood and removing waste products, and diuretics can increase the workload on these organs. Long-term use of diuretics can lead to kidney damage, impairing their ability to function properly and ultimately resulting in kidney failure . In addiStimulantsStimulants are a class of performance-enhancing substances some bodybuilders use to increase focus, alertness, and energy during their workouts. Some of the most commonly used stimulants in bodybuilding include amphetamines, caffeine, ephedrine, and clenbuterol. These substances are believed to enhance mental and physical performance, reduce fatigue, and improve endurance . While sCaffeine, found in many beverages and dietary supplements, is the most widely used stimulant in bodybuilding. While caffeine is generally safe in moderation, excessive use can lead to negative side effects, including restlessness, nervousness, and insomnia. Caffeine can also increase heart rate and blood pressure and cause gastrointestinal problems such as nausea and diarrhea . EphedriShort-term and long-term health effects of performance enhancing substance use in bodybuildingShort-Term EffectsAcne: Anabolic steroids can cause acne, particularly on the face, chest, and back. This is because steroids increase sebum production, which can clog the pores and lead to the development of acne. In severe cases, acne can become infected and lead to scarring .Baldness: Anabolic steroids can cause hair loss, particularly in men genetically predisposed to baldness. This is because steroids can increase the production of dihydrotestosterone (DHT), a hormone that causes hair follicles to shrink and eventually stop producing hair .Liver damage: Anabolic steroids can cause liver damage, including liver tumors and cysts. Recognizing that the reported liver damage, including liver tumors and cysts, is associated with such misuse is crucial. Steroids are metabolized in the liver, and prolonged or inappropriate usage can lead to liver complications. In severe cases, liver damage may necessitate a liver transplant .Cardiovascular problems: Performance-enhancing substances can increase blood pressure and cholesterol levels, leading to cardiovascular problems such as heart attacks and strokes. Anabolic steroids can also cause the heart to enlarge, which can increase the risk of heart failure and lethal arrhythmias. Additionally, using stimulants such as ephedrine can cause irregular heartbeats and increase the risk of heart attacks . It is iLong-Term EffectsKidney damage: The kidneys are responsible for filtering waste and excess fluids from the body. Prolonged use of performance-enhancing substances such as anabolic steroids, diuretics, and growth hormones can lead to kidney damage and failure. The excessive workload on the kidneys can cause them to become enlarged, leading to kidney stones and other complications .Infertility: Performance-enhancing substances, particularly anabolic steroids, can disrupt the normal production of hormones in the body, leading to infertility in both men and women. Anabolic steroid use can reduce testosterone production in men, decreasing sperm count and motility. In women, anabolic steroids can disrupt the menstrual cycle and cause infertility .Cardiovascular problems: Prolonged use of performance-enhancing substances can increase the risk of cardiovascular problems, including heart disease and stroke. Anabolic steroids can cause an increase in LDL cholesterol (the "bad" cholesterol) and a decrease in HDL cholesterol (the "good" cholesterol), which can lead to the buildup of plaque in the arteries. This can increase the risk of heart attack and stroke .Increased risk of cancer: Some performance-enhancing substances have been linked to increased cancer risk. Anabolic steroids have been linked to an increased risk of liver, kidney, and prostate neoplasms. Growth hormone has been linked to an increased risk of colon cancer . It is iThe regulation of performance-enhancing substances in bodybuildingThe Role of Regulatory agenciesRegulatory agencies, such as the WADA and the IOC, regulate performance-enhancing substances in sports, including bodybuilding. These agencies play a critical role in ensuring fair competition and protecting the health of athletes . WADA waThe IOC, on the other hand, is responsible for organizing and overseeing the Olympic Games. The IOC also works closely with WADA to ensure that athletes who compete in the Olympics comply with anti-doping regulations. The IOC has a zero-tolerance policy towards doping and works with national anti-doping organizations to enforce anti-doping regulations . In addiThe Current State of RegulationsThe current state of regulations on using performance-enhancing substances in bodybuilding is complex and varied across different countries and regions. While there are regulatory agencies that aim to control the use of these substances, such as the WADA and the U.S. Anti-Doping Agency (USADA), their efforts have not been entirely successful in eliminating the use of these substances in bodybuilding . One of Additionally, there is a lack of oversight and regulation in the manufacturing and distribution of these substances, which can lead to the production of unsafe and contaminated products. This can increase the health risks associated with their use and further complicate efforts to regulate them . OverallThe Effectiveness of Current RegulationsThe effectiveness of current regulations in curbing the use of performance-enhancing substances in bodybuilding is a topic of debate among experts in the field. While some argue that the penalties for using these substances are not severe enough to deter athletes from using them, others argue that the testing methods used by regulatory agencies are not always accurate or effective in detecting the use of these substances . In manyAnother issue with the effectiveness of current regulations is the lack of education and awareness among bodybuilders and fitness enthusiasts about the risks associated with using these substances. Many athletes are unaware of the potential health consequences of using performance-enhancing substances, and some may even believe these substances are safe to use. More must be done to educate athletes and the general public about the dangers of performance-enhancing substance use in bodybuilding and encourage safe and healthy sports practices ,48.Public health implicationsLegal Status of Performance-Enhancing Substances in Various CountriesThe use and possession of performance-enhancing substances such as anabolic steroids, growth hormones, insulin, and diuretics are regulated differently worldwide. In some countries, these substances are legal only with a prescription, while in others, they are entirely illegal and can result in severe legal consequences . For exaHowever, it's worth noting that certain countries have more lenient laws regarding the use of performance-enhancing substances. In Mexico and Thailand, anabolic steroids are legal and can be purchased over the counter without a prescription. In Brazil, anabolic steroids are legal, but their purchase requires a prescription from a licensed physician . The legRole of Healthcare Professionals in Preventing Performance-Enhancing Substance Use in BodybuildingHealthcare professionals are critical to preventing performance-enhancing substances in bodybuilding and safeguarding public health. They can educate their patients about the risks associated with these substances, including short-term and long-term health effects. Additionally, healthcare professionals can provide information on safe and effective training and nutrition strategies to help individuals achieve their fitness goals healthily and sustainably . AssessiHealthcare professionals are also well-positioned to identify individuals at risk for performance-enhancing substance use. This includes individuals with body image issues, low self-esteem, or a history of substance abuse. By addressing underlying psychological or emotional issues contributing to the desire to use these substances, healthcare professionals can provide crucial support and guidance . FurtherImpact of Performance-Enhancing Substance Use on SocietyThe impact of performance-enhancing substance use extends beyond individual health consequences and significantly affects the broader community. One major concern is that the use of these substances promotes a culture of cheating and unfair competition, eroding trust in sports and other competitive arenas. When individuals use performance-enhancing substances to gain an unfair advantage, it undermines the integrity of the competition. It diminishes the achievements of athletes who have worked hard without using such substances . MoreoveFurthermore, the pressure to use performance-enhancing substances can extend beyond individual athletes to the broader athletic community. Other athletes may feel compelled to use these substances to remain competitive, creating a dangerous substance use and abuse cycle. This culture of dependency on performance-enhancing substances can have long-lasting and far-reaching effects on the health and well-being of individuals and communities .Prevention and intervention strategiesAvailable Prevention and Intervention StrategiesPrevention and intervention strategies can effectively prevent and address the use of performance-enhancing substances in bodybuilding. One important strategy is education and awareness programs focusing on the risks associated with performance-enhancing substance use and safe and effective training and nutrition strategies. Education and awareness programs can be directed toward athletes, coaches, trainers, parents, and the general public. These programs can be delivered through various channels, including social media, websites, schools, sports clubs, and community organizations . AnotherInterventions may include counseling, behavioral therapies, and pharmacological treatments to help individuals overcome substance abuse and addiction. Counseling and behavioral therapies can help individuals address underlying issues contributing to substance abuse, such as stress, anxiety, or depression. These interventions can help individuals develop coping skills, improve communication, and strengthen social support networks. Pharmacological treatments may also help individuals manage withdrawal symptoms and cravings associated with substance abuse . FurtherImportance of Education and AwarenessEducation and awareness programs are essential to preventing the use of performance-enhancing substances in bodybuilding. These programs can give athletes the necessary knowledge to make informed decisions about their health and performance goals. By educating athletes on the potential risks and negative health consequences associated with performance-enhancing substances, they can make informed decisions that prioritize their health and well-being . MoreoveEducation and awareness programs can also promote a culture of fair competition and sportsmanship. When athletes know the risks and consequences associated with performance-enhancing substance use, they are more likely to avoid using these substances and compete on a level playing field. This can lead to a healthier and more positive sporting culture that values safe and fair competition over winning at all costs .Role of Policymakers in Addressing Performance-Enhancing Substance Use in BodybuildingThe role of policymakers in addressing performance-enhancing substance use in bodybuilding cannot be overstated. Policymakers can help ensure that performance-enhancing substances are not widely available and that access to them is limited. This can be done by developing and implementing regulations restricting these substances' sale and distribution. For example, some countries have laws requiring individuals to have a prescription before purchasing performance-enhancing substances such as anabolic steroids. Other countries have banned the sale and distribution of these substances entirely . In addiPolicymakers can also provide resources for prevention and intervention programs. These programs can educate athletes about the risks associated with performance-enhancing substances and provide information about safe and healthy ways to achieve their goals. Prevention and intervention programs can also provide support and resources for individuals struggling with substance use issues . FinallyIn conclusion, this review article has discussed the dark side of bodybuilding, specifically the use of performance-enhancing substances and their public health implications. The article has highlighted the history and evolution of performance-enhancing substance use in bodybuilding, the types of substances used, and their short-term and long-term health effects. The article has also discussed the regulation of performance-enhancing substances, the legal status of these substances in different countries, and the role of healthcare professionals and policymakers in preventing their use. Prevention and intervention strategies have also been discussed, including the importance of education and awareness, regulation, and intervention strategies. It is important to address the dark side of bodybuilding and the use of performance-enhancing substances because they pose a significant threat to public health. The use of these substances can lead to serious short- and long-term health consequences and contribute to the development of an unhealthy and dangerous culture of competition. The current state of regulations and prevention efforts is insufficient, and more must be done to address this issue. Future research in this area should focus on developing more effective prevention and intervention strategies and a better understanding of the short-term and long-term health effects of performance-enhancing substance use. Research should also explore the social and cultural factors that contribute to the use of these substances and how they can be addressed. Overall, a comprehensive and coordinated effort is needed to address the dark side of bodybuilding and promote safe and healthy practices in this sport.


The flexible high-neck flange is connected to the circular hollow section (CHS) tube through welding, and the placement of the weld seam and corresponding stress concentration factor (SCF) are crucial determinants of the joint’s fatigue performance. In this study, three hybrid models combining ant colony optimization (ACO), a genetic algorithm (GA), and grey wolf optimization (GWO) with a random forest (RF) model were developed to predict the stress distribution on the inner and outer walls of the CHS tube under different flange parameter combinations. To achieve this, an automated finite element (FE) analysis program for flexible high-neck flange joints was initially developed based on ABAQUS 2020 software. Parameter combinations were randomly selected within a reasonable range to simulate the nonlinear mechanical behavior of the joint under uniform tension, generating a dataset comprising 5417 sets of data. The accuracy of the FE model was validated through experimental data from the literature. Based on this, feature importance analysis was conducted to reveal the influence of different variable parameters on the stress distribution in the tube of the joint. The flange parameters and tube stress distribution are considered as inputs and outputs, respectively. Three hybrid RF models, specifically ant colony optimization-based random forest (ACO-RF), genetic algorithm-based random forest (GA-RF), and grey wolf optimization-based random forest (GWO-RF), are trained for regression prediction. The results demonstrate that the three hybrid models outperform the original machine learning model in predictive accuracy. The ACO-RF model achieved the highest accuracy with average coefficients of determination ( Flange connections represent a prevalent form of node connection in structural engineering . This inhttps://www.sohu.com/a/311598972_652081, accessed on 20 September 2023).Flexible flanges are typically forged in a single operation, forming both the flange plate and the flange neck, also known as flexible high-neck flanges. Under tensile loads, their characteristic of low stiffness leads to significant nonlinear mechanical behavior. This manifests as the flange plate separating under high tensile stress, generating a prying force, and inducing bending stress in the flange neck and connected tube. The widespread application of flexible high-neck flange connections is dictated by the unique requirements of different engineering projects. These distinctive features result in a wide variety of flange shapes. For instance, flange plates used in wind-turbine towers are often thick, with a substantial distance between the bolt holes and the free edge of the flange plate. These unique characteristics also give rise to variations in the nonlinear mechanical behavior of the flanges. Consequently, flange joints designed to suit specific engineering characteristics may no longer adhere to the basic assumptions of universally applicable flange engineering design methods. Therefore, instances of flange weld seam failures have been documented . This aspect has not yet received adequate attention in current structural design . Some deUnder high tensile stress, flexible flange joints exhibit nonlinear mechanical behavior, thus demanding special attention in the calculation and design process. When the tensile loading exceeds the sum of the compressive forces generated by bolt pretension, two mutually compressed flange plates will separate and open. As the tension continues to increase, the flange plates will rotate around a specific axis, and this rotation axis is different for the inner and outer flanges. Typically, the specific position of the rotation axis usually needs to be determined based on finite element analysis. Relevant design standards, such as GB50135-2017 [Furthermore, for flexible flange joints subjected to fatigue loads, targeted efforts should be made to conduct fatigue-resistant design for weld seams and bolts. Standards and specifications such as IIW  and DNVGIn summary, existing design standards are gradually becoming inadequate for the refined design of current flexible high-neck flanges. Especially for joint configurations that deviate from traditional calculation assumptions, determining rational assumptions for computational models and selecting appropriate flange design parameters to achieve an optimal design solution is an urgent issue that needs to be addressed.For addressing specialized structural designs that conventional standards may not cover, researchers typically propose corresponding theoretical calculation formulas through mechanism analysis and theoretical derivation. Alternatively, numerical simulations may be utilized to generate a large dataset of computational results, which can be used to fit semi-empirical or empirical formulas. For example, in the case of predicting SCF under different weld seam shapes, Luo et al. ,7 and WaThe era of data science has brought about a transformation in the development of various disciplines. The increase in computing capability means that previously complex problems no longer require simplified solutions. For the issue of stress distribution in flexible high-neck flange joints under tensile stress, which is influenced by numerous parameters, establishing nonlinear relationships among multiple parameters is currently one of the relatively mature techniques in the field of machine learning, especially when there exists a large dataset. As a crucial component of artificial intelligence, machine learning offers valuable methods for identifying relevant data features . In receIt is evident that applying machine learning methods to predict the stress distribution in the CHS tube of flexible high-neck flange joints is technically feasible. Based on an extensive literature review, it is known that among various machine learning models for regression prediction, the random forest model has shown the best performance in multiple cases, as seen in references ,20,21. HAt present, there are few targeted and detailed studies on the influence of different geometric parameters and load levels on the nonlinear mechanical behavior of flange joints. In addition, the research on the nonlinear mechanical behavior of flange joints focuses mostly on bolts, with few studies focusing on the stress distribution on CHS tubes connected to flanges. However, with the increasing use of high-strength steel for lightweight design in engineering applications, the thickness of the CHS tube wall is also on a downward trend, and the stress distribution near the weld seam on tube wall should also be taken into account seriously. In fact, closer to the flange plate, there are higher stress peaks on the tube. This can be attributed to the geometric asymmetry of the flange and the nonlinear mechanical behavior, resulting in a stress concentration effect. This combines with the SCF caused by the weld seam shape to form the total SCF at the weld seam location of the flange joint. For joints subjected to severe fatigue loads, determining the stress distribution on the tube is fundamental for placing weld seams and accurately assessing stress amplitudes at the weld seam location. It necessitates targeted research across a wide range of flange parameters, covering commonly used shapes of flexible flange joints. It will ultimately lead to the development of a stress distribution prediction tool that can be utilized by researchers and engineers. For applications that require user interaction and usage, a graphical user interface (GUI) is especially important. The GUI achieves a more user-friendly and intuitive stress distribution prediction tool by providing graphical elements such as icons, buttons, windows, and menus.In response to the research gaps mentioned above, this paper conducts a detailed study on the nonlinear mechanical behavior of flexible high-neck flange joints under uniform tensile load, as well as the stress distribution law on the tube wall. This paper firstly compiles a Python program to control ABAQUS 2020 software to perform automated modeling analysis and stress extraction for flexible high-neck flange joints, establishes a dataset consisting of 5417 data, and reveals the typical stress distribution of the tube wall and the influence of different geometric parameters and loads on the stress distribution. On this basis, three hybrid models, namely the ACO-RF model, GA-RF model, and GWO-RF model, were established and trained to predict the stress distribution on the tube wall of a flexible high-neck flange joint. The best performing model was selected to develop a GUI tool for engineers and researchers to predict the stress distribution in the tube wall of flexible high-neck flange joints. Finally, a case study on the fatigue damage assessment of a flexible high-neck flange joint in a wind-turbine tower is presented to demonstrate the application of the proposed model in this study.In this chapter, an overview of the random forest, ACO-based RF model, GA-based RF model, and GWO-based RF model is provided.Random forest, introduced by Leo Breiman , draws iIn the RF model, there are numerous parameters that need to be determined before training, which are known as hyperparameters. These hyperparameters include: (1) The RF model allows individual decision trees to use a maximum number of features (max_features). Increasing max_features generally enhances model performance but may reduce algorithm speed. (2) A higher number of decision trees (n_estimators) in the RF model can increase model stability but also raises the overall computational load. (3) The number of leaf nodes at the end of decision trees in the RF model (min_samples_leaf) must be considered. Smaller leaf nodes make it easier for the model to capture noise in the data. (4) The maximum depth of the decision tree in the random forest (max_depth) can limit the complexity and computation of the model. (5) The minimum number of samples needed to repartition the nodes inside the decision tree in a random forest must be considered, which limits the conditions for further partitioning of the subtree. The selection of these hyperparameters is crucial for building an RF model with the best performance and generalization ability, and therefore needs to be carefully adjusted. Different combinations of hyperparameters may apply to different problems and datasets, so trade-offs need to be made in model selection and tuning.Using ant colony optimization (ACO) for the selection of the hyperparameter of an RF model can yield stable and highly accurate regression results. The ACO algorithm was initially introduced by Marco Dorigo and his colleagues ,29,30 inThe ACO-RF hybrid model combines the advantages of both the RF model and the ACO algorithm to optimize the hyperparameters of the RF model, leading to the optimal parameter selection results. The framework of ACO-RF, as shown in the In addition to the ACO algorithm, this study also employed another optimization algorithm, the genetic algorithm (GA), to optimize the RF model. GA is a programming technique designed to create optimal solutions for any specific problem . These sThe second hybrid model and the third hybrid model used in this study, GA-RF , share aIn addition to the ACO algorithm and GA algorithm, this study also employed another optimization algorithm, grey wolf optimization (GWO), to optimize the RF model.The GWO algorithm  simulateTherefore, the third hybrid model used in this study, GWO-RF , shares To create a database for developing an RF model predicting the stress distribution of CHS tubes in flexible high-neck flange joints, a wide range of parameters has been considered, encompassing several key geometric parameters of the flange joints, as shown in 017-2017  and VDI2017-2017 .(1)8⋅ttPretension, and Load, it arises from the amalgamation of databases with different parameter ranges. However, it is crucial to emphasize that, upon examination, these non-uniformities will not affect the accuracy of the subsequently trained ML model across the entire range. Under the distribution depicted in In this study, ABAQUS 2020 software was utilized to construct the database for machine learning. A Python compiler was employed to invoke ABAQUS for modeling, analyzing, and extracting the stress results of flange joints under various combinations of parameters, as shown in tubeD/25, the stress calculation results have converged. Therefore, for subsequent parameter analysis, tubeD/25 is chosen as the mesh size. In addition, the computed results obtained by this modeling method have been compared with the experimental data from reference [eference , as showPretension), and external load size (Load). The output variables represent the stress distribution on the CHS tube wall of a flange joint under different geometric and load parameters, as shown in The performance of the RF hybrid model largely depends on the selected database input variables. In this study, a total of 10 input variables were chosen based on flange dimensions .Through training, it was determined that the hybrid models outperform the original RF model in terms of accuracy, while also eliminating the cumbersome process of manually adjusting hyperparameters. In terms of predictive performance, the ANT-RF model demonstrated the best performance. As a result, this paper developed an open-source GUI program based on the ANT-RF model for use by engineers and researchers to predict the stress distribution in the CHS tube of flexible high-neck flange joints, as shown in Under tensile stress, the flexible high-neck flange exhibits significant nonlinear mechanical behavior, resulting in bending tensile stress on the outer surface of the CHS tube. Near the flange, high stress peaks can be detected. As the distance from the flange root increases, the stress level rapidly decreases to below the nominal stress, and then returns to the nominal stress level. On the inner surface of the tube, bending compressive stress is formed under tensile stress, resulting in an opposite trend compared to the outer surface.Feature importance analysis reveals that the most influential parameter on the stress distribution in the tube of flange joint is the magnitude of the tensile force. In addition to this, the number of bolts, wall thickness of the tube, and the positioning dimensions of the bolt holes follow in terms of their respective levels of impact, each playing a non-negligible role in the stress distribution.For the specific case discussed in this study, the performance of the ACO-RF model stands out as the most outstanding among the three hybrid models with an average coefficient of determination (The following conclusions can be drawn:The tool developed in this study can compute the stress distribution on the CHS tube of the flexible flange joint under different parameters and loads. It assists in determining the optimal weld seam location and SCF. It is worth mentioning that the computed results of stress distribution can be normalized into the form of an SCF, characterizing the stress concentration caused by the macro-geometry of the flange and nonlinear mechanical behavior. This SCF, in conjunction with the SCF attributed to weld seam shape and potential misalignments, collectively form the comprehensive SCF for the weld in the flexible flange joint. This comprehensive SCF assessment provides a crucial foundation for conducting fatigue evaluations, employing local stress-life approaches such as the hot-spot stress method, effective notch stress approach, and similar techniques. In the future, building upon the research presented in this paper, further exploration will be conducted on the impact of various flange parameters on bolt stresses. This will lead to the development of a comprehensive predictive model for stress distribution in flexible high-neck flange joints, encompassing both weld seams and bolts. Subsequently, this model will serve as the foundation for assessing the competitive mechanisms of fatigue failure between weld seams and bolts in this joint configuration, as well as for conducting fatigue reliability evaluations at the joint level.


The joint analysis of single-cell transcriptomics, proteomics, lipidomics, metabolomics and spatial metabolomics is continually transforming our understanding of the mechanisms of metabolic reprogramming in tumor cells. Since head and neck tumor is the sixth most common tumor in the world, the study of the metabolic mechanism of its occurrence, development and prognosis is still undeveloped. In the past decade, this field has witnessed tremendous technological revolutions and considerable development that enables major breakthroughs to be made in the study of human tumor metabolism. In this review, a comprehensive comparison of traditional metabolomics and spatial metabolomics has been concluded, and the recent progress and challenges of the application of spatial metabolomics combined multi-omics in the research of metabolic reprogramming in tumors are reviewed. Furthermore, we also highlight the advances of spatial metabolomics in the study of metabolic mechanisms of head and neck tumors, and provide an outlook of its application prospects. The occIn this review, we mainly summarize the development from traditional metabolomics to spatial metabolomics combined with multi-omics analysis, demonstrate the comparison of the applications of different platforms of spatial metabolomics, and discuss the data-mining challenges that spatial metabolomics analysis is facing nowadays. In addition, we orderly review the novel applications of spatial metabolism combined with multi-omics analysis in the study of tumor metabolism mechanism, and also focus on its latest research in head and neck tumors. Then we end with the prospects for its application in the diagnosis and treatment of head and neck tumor metabolism.2Spatial metabolomics integrates MSI and metabolomics  technoloMSI is a new imaging method that directly scans biological samples through mass spectrometry technology. It possesses the advantages of no labeling, multipoint detection, and high sensitivity, and it can simultaneously analyze the spatial distribution characteristics of hundreds of molecules on the same tissue section. With the continuous development of modern science and technology, and meeting deeper scientific research needs, various MSI types have been developed, including matrix-assisted laser/desorption ionization mass spectrometry imaging technology (MALDI-MSI) , desorptThe different platforms of MSI and their application conditions are summarized in the following table Table 1.The metabolome is a collection of small molecular chemical entities which can involve in metabolism and maintaining the normal growth and development of organisms , and theMetabolomics is a powerful analytical tool for studying metabolite spectrum, metabolic changes, metabolic pathways, and the discovery of biomarkers, so the ultimate goal of studying metabolomics is to make a qualitative and quantitative analysis of all metabolites in a biological system as much as possible. Traditional metabolomics mainly relies on these three technical platforms: nuclear magnetic resonance (NMR) , Gas Chrin vivo  . By explSuch problems often exist in actual scientific research processes, such as many genes and proteins are obtained after the differential comparative analysis, but their function seems to be not directly related to the phenotypic differences. Therefore, the significance of spatial metabolomics combined with multi-omics analysis is also reflected here. For the hot topics of the research field, a great number of reports have revealed the direct association of phenotypes with some key metabolites. And for emerging themes of the research fields, combined multi-omics analysis can provide a more panoramic network of molecular associations. Ultimately, among all the various differential omics molecules, the interrelated functional metabolites or pathways are selected for key discussion and research, to provide more directional guidance for the pioneers in this field.3.2Lipids are important biomolecules that play multiple roles in the cellular functions of mammalian organisms. Driven by the biological significance of lipids, lipidomics  is consiin vitro research of drugs and endogenous metabolites. After finishing modeling, combined with MALDI-MSI and lipidomics, they studied the drug accumulation time and spatial distribution of hydroxychloroquine (HCQ) and its impact on lipid metabolism in lung cancer multicellular spheroids  and its potential toxicology mechanisms in breast cancer xenografts and tumor metastasis-related tissues (e.g. liver and kidney) in breast cancer xenografts treated with BPF . MoreoveASS1 are both upregulated in tumor sections. And the visualized results have shown that the expression of most lipids in gastric adenocarcinoma tissues is elevated except the expression of Phosphatidylserine (PS) and PI  and PI . It is n) and PI .Multi-omics analysis combined with spatial metabolomics can improve the integrity of biological information analysis and can visually detect gene, gene transcripts (RNA), various proteins that perform different biological functions, and even the various metabolites in the subordinating metabolic pathways. In conclusion, multi-omics analysis can help us expand our vision of the scientific field better and obtain a deeper understanding of the significance of omics interactions at different levels.4Head and neck tumors are the sixth most common tumor worldwide , with a 4.1Energy metabolism reprogramming of tumor cells promotes rapid cell growth, proliferation, and differentiation by regulating energy metabolism. And it is considered a unique marker of tumor cells , includiCYLD increases the stability and nuclear transportation of p53 by deubiquitination, to downregulate the transcription of phosphofructokinase-2/fructose-2, 6-Diphosphatase 3 (PFKFB3)   staining(PFKFB3) . Further(PFKFB3) . The rese future .4.2Early tumor diagnosis is essential to prolong patients’ survival time and improve their quality of life and prognosis . The traWith the continuous research on the mechanism of tumor metabolic reprogramming, scientists have found that metabolic reprogramming of tumor cells is often driven by a variety of signaling pathways and transcription factors . TherefoAccording to the latest application of spatial metabolomics in the diagnosis and classification of head and neck tumors, Luojiao Huang et al. have applied spatial metabolomics analysis by using AFADESI-MSI technology to thyroid tumors . Firstly4.3Tumor-targeted therapy is defined as the use of drugs that target different cancer cell targets and do not cause simultaneous harm to non-cancer cells as conventional chemotherapeutic drugs do . The purRecently, amino acid metabolic reprogramming in head and neck tumor is attracting increasing attention, especially in glutamine metabolism. Glutamine, despite being a non-essential amino acid, is the most abundant amino acid in circulation . In the de novo fatty acid synthesis to meet their special lipid needs (in vivo may depend on lysophospholipid (LP) supply. These findings have confirmed that the hypoxia-induced fatty acids uptake and lipid droplet accumulation of tumor cells constitute lipid metabolism reprogramming, which can effectively prevent the toxicity caused by ROS accumulation, thus providing higher survival benefits and treatment resistance for tumor tissues. Furthermore, it is of great significance to the research of HNSCC metabolic mechanisms and provides targets that can be further explored for the treatment of HNSCC, which has a broad clinical application prospect.Additionally, the reprogramming of lipid metabolism in head and neck tumors is also being further researched by scientists around the world. Normal body cells rely mainly on the free fatty acids from the food to synthesize the lipid components. Due to the vigorous metabolism of tumor cells, they need a large amount of lipid supply. Therefore, tumor cells depend more on id needs , thus unid needs . Schmidtid needs . They haid needs  accumulaAs an important component of personalized cancer therapy, novel drugs that target to tumor metabolism have progressed from preclinical studies to clinical trials, and in some cases have shown great efficacy. It is believed that with the advancement of drug metabolism analytical technology, researchers will be able to discover more new drug targets, construct appropriate patient models and accurately identify target patients, so as to provide patients with more drug choices and to better improve patients’ survival rate.5In 1927, Otto Heinrich Warburg discovered the Warburg effect and proposed the concept of metabolic reprogramming of tumor cells, which made the research on tumor metabolism the focus topic in tumor research today. Tumor metabolic reprogramming is one of the important features of malignancy, and aerobic glycolysis is one of the most typical features. In this review, we firstly summarized the development and challenges of spatial metabolomics, and the latest researches of spatial metabolomics combined multi-omics analysis in the metabolic mechanisms of tumor. On the basis, we reviewed the novel researches and applications of metabolic reprogramming in head and neck tumors with the technology of spatial metabolomics. Finally, we made an outlook of it to the future.Firstly, we should understand that the regulation of gene expression is one of the main sources of genetic variation, transcription predicts gene function and reveals the sequence of a gene that can be a cis-acting element, proteins are the executors of biological function, metabolism is the description of a phenotype, and morphology is the concrete presentation of regulatory results. Only through the systematic combination of genomic and phenomic data, can we truly realize the closed loop of big data for biological systems. In biological tissues, the global analysis of non-functional metabolites with spatial distribution is key to understanding biomolecular processes . The divUp to now, the unique position of spatial metabolomics in the field of tumor research has been widely recognized, and it has begun to be applied in the study of metabolic mechanisms of human tumors and new drug development. In addition to these studies mentioned above, Jian Shen et al. have evaluated the response of NSCLC patients to neoadjuvant therapy using spatial metabolomics  to judgein vivo, and to provide more visual evidence for the specific manifestation and intrinsic root of tumor metabolic reprogramming. Additionally, we are expecting to discover more sensitive biomarkers for the early diagnosis of head and neck tumors, and also to discover novel targets for drug-targeted therapy. If we can combine spatial metabolomics technology and robot surgery technology into the clinical routine workflow furtherly, through the study of a large number of clinical data to screen the appropriate tumor markers, it can be more efficient, sensitive, and accurate in early diagnosis, classification diagnosis and intraoperative margin analysis of head and neck tumors. Furthermore, it can even meaningfully instruct oncology therapy, hence better prolonging patients’ survival time and improving their long-term quality of life.At present, spatial metabolomics has shown a vigorous development trend in the study of metabolic mechanisms of human tumors, yet new applications and researches remain underdeveloped in the field of head and neck tumors. Therefore, further research is warranted. We have gained many valuable lessons from research on the metabolism of other human tumors with the help of spatial metabolomics technology, which offers important guidance to our otolaryngology head and neck surgeons to study the metabolic mechanism of head and neck tumors. Combined with IHC analysis and scRNA-seq of head and neck tumors previously conducted by our research group , we can YZ: literature search, visualization, writing - original draft preparation. CL: investigation, supervision, revised the manuscript. YC: data analysis, investigation, supervision. HD and SG: funding acquisition, project administration. ZS: idea for the review, conceptualization, writing - reviewing and editing, project administration, funding acquisition. All authors wrote one part or several parts of the manuscript. All authors contributed to conceptualising and writing the manuscript. All authors read and approved the final manuscript. All authors contributed to the article and approved the submitted version.


The COVID-19 pandemic has significantly affected various aspects of education, including the occurrence of injuries among Korean students. This study aims to analyze and compare injury rates in elementary, middle, and high schools before and after the pandemic and identify the associated factors. A non-experimental quantitative dataset compiled from the Korea School Safety Association’s annual reports (2018–2022) was utilized. The data included information on school safety accidents among Korean children and adolescents during the COVID-19 pandemic. The dataset was analyzed based on factors such as time, location, type of accident, and injured body part. The findings revealed a decline in accidents during the early phase of the pandemic, followed by an increase after schools reopened. There were notable variations in the accidents in specific locations, types, and body parts affected during the pandemic, compared with the pre-pandemic period. This study highlights the importance of continuous monitoring, implementation of safety measures, and prioritization of physical activity programs and safety education to ensure a safe learning environment. Further research is recommended to track and address evolving school accidents in response to the pandemic and its aftermath. The COVID-19 pandemic has brought about unprecedented challenges and disruptions globally, significantly impacting the education sector as well . In respWith the implementation of measures such as remote learning, intermittent school closures, and changes in school environments, it is crucial to examine the effects of the pandemic on different aspects of school life, including the occurrence of injuries in Korean elementary, middle, and high schools. Several studies have investigated the impact of the pandemic on school safety and injury rates. Injuries in schools can have various causes, including accidents during physical activities, conflicts among students, and other safety-related accidents . PandemiIntermittent school closures and their subsequent reopening have affected students’ daily routines, social interactions, and physical environments . These cThe implementation of lockdowns and related restrictions during epidemics, such as the COVID-19 pandemic, leads to substantial alterations in people’s daily routines and activities. These changes in behavior can have a direct impact on the occurrence and nature of injuries . For insAs a part of the importance of promoting overall health, the U.S. Department of Health and Human Services released “The Physical Activity Guidelines for Americans” in 2018 , which pExamining injury rates in Korean schools is very important because several unique aspects of the Korean education system precipitate injury occurrence. For instance, the Korean education system is renowned for its high academic pressure and competitiveness, which may influence students’ behavior and vulnerability to injuries. Additionally, Korean students often spend long hours studying, which may lead to potential physical strain and exhaustion and raise a concern for their overall safety . The preThis study utilized a non-experimental quantitative dataset obtained from the annual reports of the Korea School Safety Association’s file library, covering the period from 2018 to 2022, to examine the occurrence of school safety accidents among children and adolescents in Korea. The dataset provided crucial information on the number of accidents based on various factors such as time, location, type, and body part affected.schoolsafe.or.kr) (accessed on 8 May 2023). Subsequently, the school principal or parents submit a claim form and various supporting documents  related to the accident. The safety guarantee association then provides compensation and notifies the claimant of the results. Through this system, information about accidents occurring in South Korean kindergartens and elementary, middle, and high schools is all recorded in a data format. The collected data, as per the Information Disclosure Act, are published on the official website of the Central School Safety Guarantee Association on an annual basis.In South Korea, an institution called the “School Safety Guarantee Association” operates the School Safety Accident Compensation Insurance program. This program aims to provide prompt and appropriate compensation to students, school staff, and education activity participants who suffer damages due to school safety accidents, based on Article 11 of the “Law on School Safety Accident Prevention and Compensation”. All participants in kindergartens, elementary, middle, and high schools, as well as educational facilities for lifelong learning, are mandatory subscribers to this insurance. Additionally, members of foreign schools under Article 60-2 of the “Elementary and Secondary Education Act” may voluntarily subscribe to this insurance. The beneficiaries of this insurance include students, school staff, and education activity participants, and the compensation covers all life and bodily injury accidents that occur during educational activities. The procedure for handling accidents after their occurrence involves the school’s prompt reporting of the accident to the local safety guarantee association through the Accident Control System  The number of safety accidents in schools regressed during the pre-pandemic period. (2) Accidents increased during PE classes. (3) The number of accidents in sports fields and related facilities increased compared with the pre-pandemic period . SpecifiInitially, in 2022, when educational institutions transitioned back to in-person classes and resumed most educational activities in keeping with pre-pandemic norms, there was a notable regression in the number of school safety accidents. This regression can be attributed to several factors associated with the return to a more familiar learning environment and the resumption of face-to-face instruction. The change in instructional formats played a significant role in the observed increase in safety accidents. During the pandemic, schools relied heavily on remote learning and hybrid models, where students had limited in-person interaction and were more likely to be in controlled environments at home. However, with the return to traditional in-person classes, students were exposed to a different set of risks and challenges associated with physical proximity to others, such as accidental falls, collisions, or other incidents that can occur within school premises.Moreover, the restoration of regular school routines after a prolonged period of disruption could have also contributed to the increase in accidents. The transition back to pre-pandemic norms may have caused some students to feel a sense of familiarity and relaxation, which could lead to a temporary decrease in vigilance regarding safety protocols. This could include behaviors like running in hallways, not paying attention while walking, or engaging in other activities that could increase the likelihood of accidents. It is important to note that the observed increase in school safety accidents after the return to normality does not suggest that in-person instruction is inherently unsafe. Rather, it reflects the need for continued vigilance, the reinforcement of safety protocols, and ongoing efforts to create a safe and secure learning environment for students. To address this issue, educational institutions should prioritize safety education and awareness programs to remind students of the importance of following safety guidelines and being cautious in their actions. Implementing preventive measures such as maintaining clear pathways, providing safety equipment, and promoting supervision and monitoring in high-risk areas can also help mitigate the occurrence of accidents.The number of safety accidents during PE classes increased compared with those in the pre-COVID-19 period. This can be attributed to the lack of physical activity during the pandemic, as remote learning and sedentary habits became more prevalent . LimitedRegular physical activity plays a vital role in improving various aspects of health, including cardiovascular and muscular fitness, bone health, psychological well-being, cognitive function, and brain health . By incoThe number of accidents in playgrounds and auxiliary facilities (gymnasiums) increased compared with the pre-COVID-19 period. Even before the pandemic, playgrounds and auxiliary facilities had the highest number of accidents ,31. ThisThe types of accidents that increased during the COVID-19 pandemic included physical force exposure, falls, and slipping. These types of accidents showed an upward trend compared with the pre-COVID-19 period. The increase in accidents related to exposure to physical force, falls, and slipping can be attributed to several factors. During the pandemic, changes in the learning environment and restrictions on physical activity may have led to reduced physical fitness and coordination among students .The lack of regular physical exercise and decreased opportunities for movement and outdoor activities may have contributed to the higher incidence of accidents related to physical force and falls. Furthermore, the number of safety accidents involving the hands and feet increased during the COVID-19 pandemic compared with the pre-pandemic period. These accidents often manifest as minor injuries to the wrists or ankles, which are considered to be smaller areas of the body. This increase in the number of accidents can be attributed to several factors. During the pandemic, physical inactivity and sedentary lifestyles may have contributed to weakened muscles, bones, and ligaments, making people more susceptible to injuries in these smaller areas . AdditioDuring the COVID-19 pandemic, factors such as changes in school policies, parental supervision, and environmental conditions played significant roles in shaping the safety of children and adolescents, which is in agreement with previous studies, indicating that altering school policies and recreational activities to adhere to social distancing guidelines may have influenced accident rates among students . VariatiThis study has limitations that should be considered for generalization and interpretation. The current study’s primary focus on descriptive analysis poses a limitation as it may hinder the exploration of underlying causes and mechanisms behind the observed changes in injury rates. A more comprehensive approach involving deeper investigation would help in gaining a more profound understanding. Moreover, the study heavily relies on retrospective data from annual reports, which, while providing a substantial amount of information, could be susceptible to reporting biases or incompleteness. Incorporating additional data sources or employing a prospective study design could help mitigate these potential limitations and strengthen the study’s overall credibility. By doing so, the research outcomes would be more robust and reliable, allowing for a more comprehensive analysis of the factors influencing injury rates. Limited existing literature on the long-term implications of the pandemic for school safety accidents and physical activity underscores the importance of the study’s contribution in examining the effects over a significant time frame and providing valuable insights into the potential enduring impacts on students’ well-being. To gain a comprehensive understanding of school injuries and their evolution over time, future research should adopt a multifaceted approach, such as longitudinal data, cross-cultural comparisons, in-depth case studies, and comprehensive data collection. Additionally, exploring the influence of student demographics, behavior, technology use, school policies, and community involvement would inform targeted and adaptable safety interventions for creating safer learning environments.The present study can provide practical implications for informing school policies and safety measures during pandemics and beyond. First, adapting the PE curriculum to prioritize regular physical activity, even during remote learning or intermittent school closures, could help counteract the potential decline in activity levels observed during the pandemic. Second, enhancing online safety education would equip students with the knowledge to navigate digital platforms responsibly, considering the increased reliance on technology for remote learning. Prioritizing mental health support and counseling services can address the potential impact of the pandemic on students’ mental well-being, fostering emotional resilience and reducing the likelihood of risky behaviors that lead to accidents. Strengthening communication between schools and parents would facilitate collaborative efforts to ensure student safety during remote learning periods, given the significant role parental supervision may play. Integrating targeted injury prevention programs based on the study’s insights can contribute to reducing accident rates among students. Comprehensive emergency preparedness planning and addressing safety concerns during pandemics and other crises will be essential for the safety and well-being of students. By incorporating these implications, educational institutions can create a holistic approach to prioritizing student well-being and foster positive learning experiences under various circumstances. Further research is needed to explore the specific factors that contribute to the observed changes in injury rates. Examining the types and causes of accidents as well as the specific body parts affected can provide valuable insights for targeted interventions and preventive measures. Additionally, longitudinal studies that track injury rates over extended periods and compare them across different regions could enhance our understanding of the long-term effects of pandemics on school safety.By examining the overall accident status of elementary, middle, and high school students, policymakers, educators, and stakeholders can identify areas of concern and develop effective strategies and preventive measures to enhance student safety and well-being in Korean schools. It is crucial to monitor and address accident trends and patterns continuously to ensure a safe and secure learning environment for all students. The initial decline in accidents during the early phase of the pandemic was followed by an increase after the reopening of schools. These findings emphasize the need for continuous monitoring and implementation of appropriate safety measures to develop and support programs aimed at restoring and enhancing physical activity in children and adolescents. Additionally, the systematic monitoring of students by PE teachers, along with the thorough implementation of accident prevention education for all school members, is necessary. Future research should track and observe the evolving nature of accidents in schools.