The tourist experience is a core indicator of destination management for the comprehensive evaluation of destination value. Tourist experience and tourist inspiration are important concepts in the stream of research on destination marketing and management. However, these relationships remained under-explored in the extant literature. This study examined the impact of tourist experience on tourist inspiration under the moderating impact of destination familiarity. To achieve the objective of this study, data were collected online from 622 Chinese tourists. We employed partial least squares structural equation modeling (PLS-SEM) to statistically analyze the gathered data. Findings show that four types of tourist experiences, namely education, esthetics, entertainment, and escapism, significantly and positively influenced the inspired-by state of tourist inspiration, which further influenced the inspired-to-state of tourist inspiration. Destination familiarity exerted a significantly negative moderating impact on the relationship between education experience and inspired-by state of tourist inspiration. Sensitivity analysis presents that education experience was the strongest predictor of the inspired-by state followed by aesthetics, escapism, and entertainment facets of the tourist experience. Findings contribute to the theory and practice of tourism management with a robust interpretation of tourist experience, tourist inspiration, and destination familiarity to solidify the effective management of tourist destinations. Limitations and future research directions are noted. Shopping and consumption at a tourist destination is an important tourist behavior, which fuels destination revenue and revitalizes local economies, especially in the context of declining tourist inflow during destination crises, such as the COVID-19 pandemic . TouristTourist inspiration is defined as a motivational state that drives tourists to realize consumption-related new ideas . InspiraTourists are consumers of destination products and services; however, tourist purchase decision might be different from ordinary consumer purchase decision. As a classic theory of the consumer decision process, the Engel-Kollat-Blackwell (EKB) model assumes that consumer decision-making is a completely rational process  that pasThe tourist experience is defined as an interaction between a tourist and a destination . An indiDestination familiarity means tourists’ subjective assessment of their existing knowledge and information alongside learning new knowledge and skills during visits to tourist destinations . Some toAn individual’s familiarity affects his/her information search behavior . While tThis study aims to examine the following: (1) How does tourist inspired-by state impact tourist inspired-to state? (2) How does tourist experience impact tourist inspiration, i.e., tourist inspired-by state? (3) How does tourist destination familiarity exert its moderating impact on the relationship between tourist experience and tourist inspiration, i.e., tourist inspired-by state? Findings unravel the psychological mechanism of tourist purchase motivation from the perspective of tourist inspiration. This study fills theoretical gaps with a proposed conceptual framework and offers guidelines to destination marketing organizations (DMOs) in solidifying destination management and promotion efforts to skyrocket sales revenue of tourist destination. This study provides roadmaps for scholars and practitioners to conduct future research on destination marketing and management.Customer inspiration is defined as a state of temporary motivation evoked by corporate marketing efforts, promoting the generation of new ideas related to consumption, and driving consumers to take action on new ideas . As an iExternal stimuli, such as destination environment, destination events, and discovery of new possibilities in a tourist destination, may trigger tourist inspired-by-state in tandem with generating new ideas regarding tourist consumption with self-transcendence . AccordiHypothesis H1: Tourist inspired-by state exerts a significantly positive impact on the inspired-to state of tourist inspiration.The tourist experience is the application of the experience economy in a tourism context. The concept of the experience economy is widely accepted by tourism scholars and practitioners. According to the two dimensions of customer connections, namely absorption and immersion, and the level of customer participation, namely passive and active participation, four experience realms of tourist experience are identified in the extant literature for business promotion, i.e., education, esthetics, entertainment, and escapism . A uniquEducation experience may impact tourist inspiration . TouristHypothesis H2: Education experience exerts a significantly positive impact on the inspired-by state of tourist inspiration.Personal experience linked to entertainment is the most emphasized dimension of the tourist experience in destination marketing . DestinaHypothesis H3: Entertainment experience exerts a significantly positive impact on the inspired-by state of tourist inspiration.The esthetics experience is a process in which tourists feel and appreciate objective matters and the environment . TouristHypothesis H4: Esthetics experience exerts a significantly positive impact on the inspired-by state of tourist inspiration.Escaping reality is an important motivation for tourists . TouristHypothesis H5: The escapism experience exerts a significantly positive impact on the inspired-by state of tourist inspiration.As an important construct in the field of marketing, familiarity with a product or brand refers to consumer experience and knowledge about a product or brand that may influence consumer decision-making regarding a product or brand . PreviouExisting studies show that destination familiarity has an important impact on tourist information search behavior . TouristHypothesis H6: Destination familiarity exerts a significant moderating impact on the relationship between education experience and the inspired-by state of tourist inspiration such that the relationship is weak when destination familiarity is high.Hypothesis H7: Destination familiarity exerts a significant moderating impact on the relationship between entertainment experience and the inspired-by state of tourist inspiration such that the relationship is weak when destination familiarity is high.Hypothesis H8: Destination familiarity exerts a significant moderating impact on the relationship between esthetics experience and the inspired-by state of tourist inspiration such that the relationship is weak when destination familiarity is high.Hypothesis H9: Destination familiarity exerts a significant moderating impact on the relationship between escapism experience and the inspired-by state of tourist inspiration such that the relationship is weak when destination familiarity is high.The proposed theoretical associations among the constructs of this study are presented in A survey questionnaire was designed to gather data from Chinese respondents having tourist experience. Screening questions were made a part of the survey questionnaire: (1) What type of destination was your last trip? (2) What is the name of your recently visited tourist destination? (3) How recent is your tourist experience? To measure the proposed constructs in this study, scale items were adapted from the previous studies. Scale items for education, esthetics, entertainment, and escapism dimensions of tourist experience were adapted from 1, a professional online academic survey platform in China, between November 5, 2019 and November 10, 2019, to gather data from Chinese tourists who were at least 18 years old to ensure the consent requirement of the study respondents  was employed to test the proposed relationships among tourist experience, tourist inspiration, and destination familiarity. PLS-SEM is preferred to the co-variance-based approach because PLS-SEM is one of the most used methods in analyzing the structural relationships of latent variables  and modeThe convergent validity and discriminant validity of each construct were evaluated. The average variance extracted (AVE) of all constructs ranged from 0.582 to 0.772 see , which iR2 values of the dependent variables are over 0.1 and Q2 values are over 0, presenting the predictive ability and predictive relevance of the structural model . Thus, hypothesis H1 is supported. Findings for education experience , entertainment experience , esthetics experience , and escapism experience  show that education, entertainment, esthetics, and escapism dimensions of tourist experience exert significantly positive impact on the inspired-by state of tourist inspiration. Therefore, hypotheses H2, H3, H4, and H5 are supported. Findings show that destination familiarity has a significantly negative impact on the relationship between education experience and inspired-by state of tourist inspiration , indicating that when tourist destination familiarity is high, the relationship between education experience and the inspired-by state of tourist inspiration is weak. This supports hypothesis H6. However, findings show that destination familiarity has no moderating impact on the relationship between entertainment experience and inspired-by state , between esthetics experience and inspired-by state , and between escapism experience and inspired-by state . Thus, hypotheses H7, H8, and H9 are not supported. The structural model of the study is presented in Before the bootstrapping procedure, destination familiarity was set as a moderator, inspired-by state as a dependent variable, and education, entertainment, esthetics, and escapism dimensions of tourist experience as independent variables to generate four moderating impacts. Since destination familiarity is a reflective-reflective construct in this study, a product indicator calculation method was selected while generating the moderating impact on the relationship between dimensions of tourist experience and the inspired-by state of tourist inspiration. To exclude the influence of other factors, demographic and destination variables mentioned in al model . To solial model  show thaThe objective of sensitivity analysis is to determine how much of the change in the dependent variable is caused by the change in the relevant independent variable  and to oTo analyze the importance of education, entertainment, esthetics, and escapism dimensions of tourist experience and their associated impact on the inspired-by state of tourist inspiration, we constructed one ANN model. In the ANN model, the four dimensions of tourist experience, i.e., education, entertainment, esthetics, and escapism, are in the input layer, the inspired-by state of tourist inspiration is in the output layer, and there are three hidden nodes in the hidden layer see . Table 6The findings of this study show that tourist inspired-by state significantly and positively influenced tourist inspired-to state which is in line with the previous studies demonstrating a significant and positive correlation between the inspired by state and the inspired-to state of customer inspiration . For exaThe findings of this study show that the education experience positively and significantly influenced the inspired-by state of tourist inspiration. A strong beta (β) value for the relationship between education experience and inspired-by state of tourist inspiration reflects a strong impact of education experience on the inspired-by state as compared to the impacts of entertainment, esthetic, and escapism dimensions of the tourist experience. These findings are consistent with the work of The findings of this study also show that the entertainment experience significantly and positively influenced the inspired-by state of tourist inspiration. Compared with the other three dimensions of tourist experience, entertainment experience has a weak relationship with the inspired-by state of tourist experience due to the smallest path coefficient. The impact of the entertainment experience on tourist inspiration is investigated in this study which remained under-explored in the previous studies. Destination performances and activities inspire tourists by provoking tourists’ positive emotions and feelings of happiness, which are the prominent features of the entertainment experience . From thThis study demonstrates that esthetics experience significantly and positively influenced the inspired-by state of tourist inspiration. Compared with education experience and escape experience, the path coefficient of the influence of esthetics experience on the inspired-by state is weak. These findings are consistent with the work of This study revealed that the escapism experience significantly and positively influenced the inspired-by state of tourist inspiration, which is also consistent with the work of The findings of this study revealed a negative, however, weak, moderating impact of destination familiarity on the relationship between education experience and the inspired-by state of tourist inspiration. Nevertheless, the impact of entertainment, esthetics, and escapism dimensions of tourist experience on the inspired-by state of tourist inspiration was not negatively moderated by destination familiarity and the moderating impact remained comparatively weak due to co-efficient values near zero. Destination familiarity is described as tourist cognition and tourist knowledge and experience regarding tourist destination . As tourThis study makes several important theoretical contributions to the extant literature. First, from the general perspective of tourism, this study puts forth empirical evidence to demonstrate the impact of tourist experience, i.e., education, esthetics, entertainment, and escapism, on tourist inspiration, i.e., the inspired-by state, that provides a broader lens to DMOs to understand tourist cognitive responses for destination promotion, which previously remained context-specific and limited, such as wellness tourism  and inteDestination marketing organizations can design and arrange destination performances and events to enhance tourist experience, which may motivate tourist inspiration and increase destination sales revenue. Although the discussed four dimensions of tourist experiences significantly and positively affect tourist inspiration, DMOs need to combine their advantages to strategically position tourist experience of destinations. Based on such a positioning strategy, DMOs can plan related marketing activities to highlight specific tourist experiences in tourist destination to strengthen destination brand image. For example, the southern Sichuan Bamboo Sea is in Yibin City, Sichuan Province, China, and is a 4A-level natural scenic spot in China. To highlight the educational experience and aesthetic experience, the scenic spot has built the largest bamboo professional museum in China by combining the advantages of rich bamboo resources to show tourists the long history of Chinese bamboo culture and various bamboo crafts. Visiting the museum may inspire tourists to purchase bamboo craft products to increase scenic spot sales revenue. To highlight the escape and entertainment experiences, the scenic spot plans bamboo raft water experience activities. This inspires tourists to consume bamboo raft experience projects and, ultimately, to increase scenic spot sales revenue. Thus, tourist inspiration uncovered under the empirical lens of this study provides new strategic directions for DMOs.For destinations characterized by education experience, DMOs need to consider the differences in tourist destination familiarity between revisiting and first-time tourists and adopt different marketing strategies for different types of tourists. For revisiting tourists, DMOs can use two marketing strategies to stop declining purchase motivation caused by the negative moderating effect of destination familiarity to maintain destination sales revenue. The first strategy is that using sales promotions and designing creative marketing campaigns may improve tourist purchase motivation and tourist inspiration, respectively. The second strategy is to increase tourist novelty experience during tourist visits to destinations to trigger tourist inspiration, which weakens the negative moderating effect of destination familiarity, with the help of continuous innovation in destination performances and events.COVID-19 pandemic may negatively impact tourist willingness to visit familiar and unfamiliar tourist destinations, resulting in a huge impact on the global tourism industry due to the sharp drop in the number of tourists . The COVThis study explores the relationship between the dimensions of tourist experience and tourist inspiration. However, there are some limitations of this study that open doors for future research on the topic under investigation. There are many antecedents to tourist inspiration . HoweverSince this study explored the inspired-to state of tourist inspiration with a focus on purchase motivation, the proposed theoretical framework of the study can be extended to reflect how tourist inspiration affects tourist revisit intention, tourist wellbeing, and tourist intention to recommend a destination . Future All datasets generated for this study are included in the article/This study was carried out in accordance with the recommendations of the Local Ethics Committee of Shenzhen. All the study participants provided written informed consent in accordance with the Declaration of Helsinki. The study protocol was approved by the Local Ethics Committee of Shenzhen University.JX: conceptualization, conduct of the survey, data gathering, data analysis, revisions, development, and proofreading of the manuscript. ZZ: conceptualization and survey design. SM: revisions, development, and proofreading of the manuscript. RC: data analysis. NZ: survey design. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Nanozymes are synthetic nanoparticulate materials that mimic the biological activities of enzymes by virtue of their surface chemistry. Enzymes catalyze biological reactions with a very high degree of specificity. Examples include the horseradish peroxidase, lactate, glucose, and cholesterol oxidases. For this reason, many industrial uses of enzymes outside their natural environments have been developed. Similar to enzymes, many industrial applications of nanozymes have been developed and used. Unlike the enzymes, however, nanozymes are cost-effectively prepared, purified, stored, and reproducibly and repeatedly used for long periods of time. The detection and identification of pathogens is among some of the reported applications of nanozymes. Three of the methodologic milestones in the evolution of pathogen detection and identification include the incubation and growth, immunoassays and the polymerase chain reaction (PCR) strategies. Although advances in the history of pathogen detection and identification have given rise to novel methods and devices, these are still short of the response speed, accuracy and cost required for point-of-care use. Debuting recently, nanozymology offers significant improvements in the six methodological indicators that are proposed as being key in this review, including simplicity, sensitivity, speed of response, cost, reliability, and durability of the immunoassays and PCR strategies. This review will focus on the applications of nanozymes in the detection and identification of pathogens in samples obtained from foods, natural, and clinical sources. It will highlight the impact of nanozymes in the enzyme-linked immunosorbent and PCR strategies by discussing the mechanistic improvements and the role of the design and architecture of the nanozyme nanoconjugates. Because of their contribution to world health burden, the three most important pathogens that will be considered include viruses, bacteria and fungi. Although not quite seen as pathogens, the review will also consider the detection of cancer cells and helminth parasites. The review leaves very little doubt that nanozymology has introduced remarkable advances in enzyme-linked immunosorbent assays and PCR strategies for detecting these five classes of pathogens. However, a gap still exists in the application of nanozymes to detect and identify fungal pathogens directly, although indirect strategies in which nanozymes are used have been reported. From a mechanistic point of view, the nanozyme technology transfer to laboratory research methods in PCR and enzyme-linked immunosorbent assay studies, and the point-of-care devices such as electronic biosensors and lateral flow detection strips, that is currently taking place, is most likely to give rise to no small revolution in each of the six methodological indicators for pathogen detection and identification. While the evidence of widespread research reports, clinical trials and point-of-care device patents support this view, the gaps that still exist point to a need for more basic research studies to be conducted on the applications of nanozymology in pathogen detection and identification. The multidisciplinary nature of the research on the application of nanozymes in the detection and identification of pathogens requires chemists and physicists for the design, fabrication, and characterization of nanozymes; microbiologists for the design, testing and analysis of the methodologies, and clinicians or clinical researchers for the evaluation of the methodologies and devices in the clinic. Many reports have also implicated required skills in mathematical modelling, and electronic engineering. While the review will conclude with a synopsis of the impact of nanozymology on the detection and identification of viruses, bacteria, fungi, cancer cells, and helminths, it will also point out opportunities that exist in basic research as well as opportunities for innovation aimed at novel laboratory methodologies and devices. In this regard there is no doubt that there are numerous unexplored research areas in the application of nanozymes for the detection of pathogens. For example, most research on the applications of nanozymes for the detection and identification of fungi is so far limited only to the detection of mycotoxins and other chemical compounds associated with fungal infection. Therefore, there is scope for exploration of the application of nanozymes in the direct detection of fungi in foods, especially in the agricultural production thereof. Many fungal species found in seeds severely compromise their use by inactivating the germination thereof. Fungi also produce mycotoxins that can severely compromise the health of humans if consumed. Biomedical applications of nanoparticles are currently undergoing an exponential increase ,2,3. TheUnlike prions, viral infections are ubiquitous with more than 200 viruses which are known to be able to infect and cause diseases in humans ,25. In tEscherichia coli causes food poisoning, while Helicobacter pylori and Neisseria gonorrhoeae cause gastritis and the sexually transmitted gonorrhea, respectively. The structures of bacteria are diverse, including small spherical, spiral threaded, cylindrical, flagellated rod-like, and filamentous chain-like structures. They are prokaryotes, and therefore lack well-defined nuclei and membrane-bound organelles, but have chromosomes composed of a single closed DNA circle. Virulent bacteria are classified as Gram positive or negative [Similar to viruses, bacteria are ubiquitous as free-living and mostly unicellular organisms that can aggregate to form colonies. Whereas most of the known bacteria live normally on various parts of the human body without causing illness, some bacterial species are pathogenic. For example, negative ,29. GramCoccidioides immitis, Histoplasma capsulatum, and Blastomyces dermatitidis, which account for many infections each year. Even though some fungal species are normally avirulent in healthy people, they can be transmitted to deep tissues and cause serious infections in patients with compromised immune systems. Examples include Candida albicans, Aspergillus, Rhizopus, and Fusarium species [Fungi often infect the human body through several mechanisms. A virulent fungal species can invade the body and cause disease on the skin, and then spread to other tissues and organs, such as bones, hair, and nails or affect the entire body. Examples of virulent fungal species include  species . The gen species . The bacAscaris lumbricoides, trematodes, cestodes, and nematodes. The use of nanotechnology in identifying and treating helminth infection has been widely reported [Nowadays the identification and diagnosis of several multicellular organisms which cause disease through infection as parasites can be facilitated through the use of nanotechnology. Parasitic organisms that infect humans which can be identified and diagnosed using nanotechnology include protozoa, helminths, and ectoparasites. Microscopic, single-celled protozoan parasites multiply in humans, and this contributes to their virulence and the serious illnesses that develop from infection even by a single cell. The multicellular helminths on the other hand are generally visible to the naked eye in their adult stages. Examples include worm-like creatures such as reported .One of the fastest growing applications of nanotechnology today is the identification of pathogens and diagnosis of the diseases they cause, called nanozymology, which is an emerging terminology used to refer to the science of nanozymes . NanozymBacillus subtilis, Streptococcus mutans, Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli, and Salmonella enterica. Clearly, Liu et al., (2021) have shown that nanozymology is a fast-growing science in the detection and identification of viral and bacterial pathogens that are known to cause serious illnesses among humans. This review will focus on the details of the methodologies used in this regard, highlight the innovative structure and architecture of the nanoconjugates used as nanozymes, and suggest possible further innovations on the basis of the reported mechanisms and the performance of the nanozymes.The current review will focus on the applications of nanozymes for the detection and identification of pathogenic microorganisms and diagnosis of the infectious diseases they cause. Several metal and metal oxide nanozymes have been used for the detection and identification of many microorganisms responsible for infectious diseases. Examples of methods cited by Liu et al., (2021), used for the detection and identification of microorganisms responsible for infectious diseases, include the colorimetric immunoassay, magnetic nanozyme-linked immunosorbent, enzyme-linked immunosorbent, immunochromatographic, colorimetric and electrochemical, nanozyme strip, magnetophoretic chromatography, and the lateral flow essays. Viral pathogens that have been identified and detected using these methods include the murine noroviruses, the viruses that cause the common influenza, the avian influenza, mumps, measles, and listeriosis. Bacterial pathogens include The ubiquitous distribution of virulent pathogenic microorganisms is a major cause of the infections that cause morbidity and mortality across the world. Given that environmental detection has taken the center stage in recent times due to the heightened recognition of the impact of environmental pathogenesis on epidemiology, identification of virulent pathogens in the environment before they cause infection may become more important in the future than the identification for purposes of disease diagnosis. In the developing world, pathogens in the environment are a major driver of epidemics such as cholera  and dyseThe screening of animal vectors such as vermin, pests, and pets and their environments is an essential aspect of the detection and identification of virulent pathogens. For example, much research has supported the zoonotic origins of SARS-CoV-2 ,46, and Several methods are used for the detection of pathogens. Most of these methods are specific for a limited scope of pathogens and are not usable outside their pathogen specificity scope. For example, the enzyme-linked immunosorbent assay is used to detect antibodies of the pathogens that cause specific viral and bacterial diseases ,56. It iIn the same way that the lateral flow immunoassays mimic the enzyme-linked immunosorbent assays on the lateral flow flat pad along which the analyte moves by osmosis, picking up the specific detection antibodies and the signal amplification reagents ,58, so tTwo of the most widely used enzyme-linked immunosorbent assay tactical approaches include the direct and sandwich strategies. In the direct approach, the antigen and the detection antibody are conjugated directly to form the antigen–antibody conjugate. In the sandwich approach, one antibody is first used to capture the antigen, and this is followed by conjugation with the detection antibody to form an antibody–antigen–antibody sandwich. The detection antibody is always tagged with the detection enzyme. In the case of nanozymology approaches, the detection antibody is always tagged with the detection nanozyme. In some of these studies the nanozymes are augmented with biological material such as enzymes, DNA and RNA strands or fragments ,62. HoweStaphylococcus aureus, using a bovine serum albumin capped cobalt oxide nanozyme, by attaching the pathogen to the nanozyme using a novel specific fusion phage protein, and isolating the nanozyme–pathogen complex, using magnetophoretic chromatography in an external magnetic field [Staphylococcus aureus in milk was achieved.Recently, Oh et al., (2020) reported an ultra-sensitive innovation to improve on immunological approaches by using the initial magnetic separation and concentration of the pathogen, followed by nanozyme immunoassay . In theiic field . Ultra-sDue to the multi-stage value chain, from the farm to retail sales, along which the vast majority of food contamination with virulent pathogens often occurs, foods have great potential to become sources of virulent pathogenic infections to humans . TraditiA traditional approach for the detection and identification of bacteria in foods may be illustrated with the real-time PCR detection of 12 common food-borne bacterial pathogens reported by Liu et al., (2019). Their procedure consisted of a 95 °C denaturation (30 min), 95 °C amplification (5 s × 40 cycles), 55 °C (10 s), and 72 °C (30 s) . WhereasResponding to rising fungal infections and the need for fast and reliable detection and identification of the responsible pathogens, Wagner et al., (2018) reported a comparative evaluation of the PCR analysis of the small ribosomal unit DNA, the internal subscriber region of the ribosomal DNA, and the culture method. Using the same PCR parameters, they found a high degree of congruence between the three procedures although the PCR methods were significantly faster and more reliable . The resRecently developed nanozyme strategies for detecting and identifying foodborne viral pathogens have presented an opportunity for developing ultrafast and cost-effective assays, taking the limits of detection much lower than those of the culture-and-grow, the traditional immunoassays, and the traditional PCR strategies. Although sensitively and reliably detected using the reverse transcription quantitative PCR methods, Weerathunge et al., (2019) proposed a novel colorimetric nanozyme strategy for fast and ultrasensitive detection and identification of the foodborne human norovirus responsible for gastroenteritis outbreaks worldwide. The approach combines the nanozyme activity of AG3-aptamer conjugated gold nanoparticles with the high binding specificity of the aptamer for the surrogate human norovirus, to develop a blue color producing sensor in response to this human norovirus surrogate, in which the blue color intensity is directly proportional to the virus concentration . The seqThe gold nanoparticle nanozyme activity produces an intense blue color upon interaction with the human norovirus. The AG3-aptamer conjugated nanozyme loses this property because the gold nanoparticles are concealed under the shell of the AG3-aptamer. The surrogate human norovirus, however, replaces the aptamer shell, thus restoring the blue color production activity of the nanozyme in a norovirus concentration linearly dependent manner, thus creating an ultrasensitive sensor for the norovirus. Other pathogens do not bind to the human norovirus specific aptamer. Therefore, the sensor is highly specific for the human norovirus.Yersinia enterecolitica, which is responsible for the symptoms of yersiniosis. The immunosensor detected Yersinia enterecolitica in milk to the ultralow detection limit of 5 CFU/mL [Yersinia enterecolitica immobilized on the electrode inhibits the nanozyme redox current, and this means that the detection signal is inversely proportional to the bacterial analyte concentration in the sample. The preparation of the working electrode is shown in A novel graphene quantum dot (GQD) nanozyme immunosensor strategy was reported as part of an electrochemical sensor that was developed as an efficient method for the detection and identification of the virulent bacterial pathogen 5 CFU/mL . The golGibberella zeae, has been reported [While literature is lacking on nanozyme strategies for direct fungal detection and identification, similar to natural enzymes, nanozymes have been implicated in the strategies for the destruction of fungal pathogens . Given treported . The gol2 in the place of the traditionally used enzymes, was developed for the detection and identification of aflatoxin B1 [2 which was used to mimic the peroxidase enzyme activity in this study, is a metal-organic framework that is made up of ferrous ions linked with 2-aminobenzene dicarboxylic acid [Aspergillus species in foods by a metal-organic framework-linked immunosorbent assay. The results of the study showed a 90% sensitivity and a 96% specificity which was an improvement on the measured 63% sensitivity and 90% specificity of the enzyme-linked immunosorbent assay used for comparison in the study.Although they cannot really be described as nanozymes, metal-organic frameworks, which are going through phenomenal growth in research studies and applications, also mimic a number of the same enzymes that nanozymes are known for mimicking. For this reason, they are also used in the same nanozyme-based bio-analytical procedures as nanozymes, notably for the detection and identification of pathogens. A variant of the enzyme-linked immunosorbent assay that uses Fe-MIL-88B–NHtoxin B1 . Fe-MIL-lic acid . The aflAlthough cancer cells are not explicitly included in the strict definition of pathogens, once they are formed in the body, they cause cancerous tumors, and they spread across the body as part of cancerous tumor metastasis. The formation of cancer cells including the metastasis processes are included in what is commonly referred to as cancer pathogenesis ,92. For One of these methods is a direct detection and identification of cancer cells using graphene oxide decorated with nanoparticles of platinum which are functionalized with folic acid . The metAdditionally, a copper oxide nanozyme supported on a nanocomposite consisting of graphene oxide decorated with gold nanoparticles was used to amplify the electrochemical signal for the detection and identification down to 27 circulating MCF-7 cells using cyclic voltammetry . The cycLeishmania parasites using a chronoamperometric procedure in which nanogold is deposited on the working electrode to act as the catalyst for the conversion of hydrogen ions to hydrogen, while monitoring the faradaic current as the detection signal, has been reported [Leishmania parasites.The detection of parasites appears to be mainly dependent on the indirect methodologies such as the detection of parasite ova in the water using techniques such as (i) graphic enumeration using a microscope, (ii) PCR-based techniques, and (iii) flow cytometry . These treported . This apLeishmania parasite concentration over a 7-log range. In addition, a nanogold-based enzyme-linked immunosorbent assay was used for the detection and identification of Trichinella spiralis, in which the horse radish peroxidase enzyme-linked immunosorbent assay was used alongside a nanozyme-linked immunosorbent assay, wherein the horse radish peroxidase enzyme was replaced with gold nanoparticles as the nanozyme, and polyclonal antibody immunoglobulins were used to capture and immobilize the Trichinella spiralis [The pellet obtained from the incubation of casein capped nanogold and centrifuge isolation is analyzed using a chronoamperometric procedure on a screen-printed carbon electrode. This procedure proved to be sensitive down to a single parasite per milliliter, with a linear dependence on spiralis .Eschericia coli very sensitively by efficiently mimicking the peroxidase enzyme activity, employing the sandwich type immunoassay strategy [Eschericia coli compared to other bacteria, with remarkable sensitivity down to 10 bacteria per milliliter.The nanozyme platform shown in strategy . In thisBiosensors for pathogen detection and identification vary in complexity, sensitivity, speed of response, cost, reliability, and durability, which are considered to be among the most desirable indicators. Colorimetric paper-based biosensors rival most in this regard especially when coupled with color amplification strategies . The thrSalmonella typhimurium with high sensitivity is found in a recent report on a sensor based on polymer nanospheres decorated with platinum quantum dots [Salmonella typhimurium specific capture antibodies immobilized on magnetic beads are initially used to enrich the analyte magnetically before conjugation with the detection antibodies that are tagged with platinum quantum dot decorated polymer nanospheres. The platinum quantum dot decorated polymer nanosphere nanozyme mimics the peroxidase conversion of the TMB to form its blue colored single electron oxidation product visible to the naked eye in a highly Salmonella typhimurium specific, sensitive, and pathogen analyte linearly dependent manner. This mechanism is shown in The performance of many pathogen biosensor devices can be substantially improved by using nanozymes in their biosensor mechanisms. Nanozymes can be used to detect pathogens directly by mechanisms that involve the direct interaction of nanozymes with the pathogen, or indirectly by mechanisms that involve interaction of nanozymes with chemical compounds produced by the pathogen. For example, the indirect detection of bacteria in urine will surely be improved by incorporation of the results of the many reports on rapid and selective detection of nitrite, based on nanozymes because the nitrite is one of the products of bacterial infection ,110. Thetum dots , which uAnother example appearing in a recent review is the use of photothermal immunoassay signal transduction . In thisMicrofluidic biosensors hold great promise for the detection and identification of pathogens at the point-of-care, for general and bio-analytical research, and biomedical engineering research laboratories because of the possibilities offered by the multiple channel miniature fluidic flows. A microfluidic system consists of multiple microfluid flow channels to enable virtually any of the operational steps that can be rationally imagined and designed into chemical and biological analytical procedures to take place in a miniature chip that can be as small as the tip of a finger. Microfluidic chips are constructed by elaborate carving of the microfluid flow channels, microvalves, and micro-fluid flow pumps . The micSalmonella analyte concentration, based on the color development of the nanozyme-based peroxide oxidation of TMB. The immuno-magnetic nanoparticles formed by mixing of capture antibodies with the magnetic nanoparticles followed by addition of the Salmonella sample are treated with immuno-nanozyme conjugate obtained by the detection antibodies and the manganese dioxide nanoflowers. The foregoing magnetic immunoassay conjugate reagents are thoroughly mixed in the spiral micromixer microfluid incubation chip and then magnetically captured in the separation chamber of the microfluid chip as shown in In recent times, microfluidic systems have been advanced by incorporation of nanozymology into their design and analytical strategies for the detection and identification of pathogens in food, environmental and clinical samples, affording timely analytical results. Firstly, a colorimetric microfluidic biosensor methodology was reported recently by Xue et al., (2021) which utilizes the magnetic analyte enrichment sandwich immunoassay strategy and measurement of Salmonella analyte concentration data collected from the detection chamber of the chip are captured, processed, and displayed on a smartphone [Addition of TMB causes the colorimetric reaction catalyzed by the terminal manganese nanoflowers on the magnetically captured sandwich conjugate to give the colored single electron oxidation product. In this microfluidic immunoassay strategy, the artphone .Salmonella specific monoclonal antibody conjugated immuno-magnetic beads, which capture the bacteria to form the sandwich conjugate with the Salmonella analyte. In this study, the amino-modified manganese dioxide nanozyme is conjugated with carboxyl-modified fluorescent quantum dots as the fluorescence signal generators via the 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide. The analyte, immuno-magnetic beads, and the immuno-nanozyme conjugate are injected together into the microfluidic chip for mixing and formation of the nanoconjugate consisting of the Salmonella specific monoclonal antibody immunomagnetic beads, the Salmonella, and the fluorescent quantum dot decorated polyclonal antibody immuno-nanozyme. This is achieved by thorough mixing and incubation in the microfluidic chip. After release of the quantum dots by glutathione dissolution of the manganese dioxide nanoflowers, the fluorescence of the quantum dots is measured to quantify the Salmonella. This fluorescence microfluidic biosensor methodology was reported to achieve a detection limit of 43 colony forming Salmonella per milliliter, from chicken meat derived samples. The microfluidic chips of these colorimetric and fluorescence microfluidic biosensors are illustrated in In the second example, a fluorescence microfluidic biosensor methodology was reported by Hao et al., (2020) which also utilizes the magnetic analyte immobilization sandwich immunoassay strategy, but the measurement of the analyte concentration is based on the fluorescence intensity of the released quantum dots . The immEbola, a deadly pathogen that is raging in West Africa without cure [The attractiveness of paper strip tests is their simplicity, response speed, low cost, and ease of use, characteristics that make them the preferred point-of-care test methods in such resource constrained environments such as the developing world, especially in their rural areas. A nanozyme strip was developed for rapid local diagnosis of out cure . The indout cure . It is cEbola capture antibody 1H3 in borate buffer and a control line consisting of goat anti-mouse IgG antibody in borate buffer were dispensed on an absorbent pad terminated nitrocellulose membrane strip as shown in A test line consisting of the The rate of movement of water by diffusion across the nitrocellulose membrane used in paper-based dipstick methods is generally inversely proportional to the density of the membrane. With this in mind, the variable pressure pressing of the nitrocellulose membrane introduced for the first time by Park et al., (2016) in paper-based dipstick research methodology, achieved a variable fluid flow rate due to the different density regions created on the nitrocellulose paper membrane, and demonstrated the intended impact on the sequential multi-step immunoassay reactions that take place at predetermined positions, where the various immunoassay reagents are immobilized on the nitrocellulose membrane paper strip . For thiIn this research, the sandwich type immunoassay methodology was used by locating the capture antibodies at the test line after the nanozyme tagged detection antibodies in the water diffusion direction, so that the water hydrates and mobilizes the nanozyme tagged antibodies with the pathogen, before binding the capture antibody at the test line, at which stage the desired pathogen specific nanogold tagged sandwich antibody–pathogen–antibody conjugate is formed. When the pathogen specific nanogold tagged antibody conjugate reaches the test line, the nanogold generates the amplified color signal, which is visible to the naked eye, thus enabling detection and identification of each pathogen in its specific channel test line. The slow flow channels 1 and 3 of the paper strip enable amplification of the test line colorimetric signal by generating gold ions which attach on the gold nanoparticles attached to the detection antibodies of the nanogold tagged sandwich antibody–pathogen–antibody conjugate. The paper strip working principle is illustrated in Escherichia coli and Salmonella typhimurium to the detection limits of 105 and 106 CFU/mL, respectively.Although the detection limits of the paper strip were still very high and the plan was to further reduce them by optimization of the methodology, the current value proposition of the multiple flow rate and multiple channel colorimetric paper strip reported for the first time was the simultaneous analysis of two different microorganisms. This was demonstrated by the simultaneous analysis of Salmonella typhimurium and Escherichia coli by the naked eye, much like the nanogold based nanozyme-linked immunoassay paper strips, colorimetric chemiluminescence paper strips present an elegant way to amplify the naked eye visualization of the colorimetric immunoassay detection signal. A paper strip based on an iron oxide magnetic nanoparticle nanozyme-linked immunoassay significantly enhanced the detection and identification of the SARS-CoV-2 spike protein receptor-binding domain derived antigen by visualizing the chemiluminescence signal obtained from nanozyme catalytic conversion of luminol [Whereas the different flow rate multiple channels amplified colorimetric nanogold paper strip reported by Park et al., (2016) enhanced the nanogold signal to achieve superior detection and identification of  luminol . In thisA sandwich type immunoassay method is used. The sample containing the SARS-CoV-2 spike protein derived receptor-binding domain antigen binds with the nanozyme tagged capture antibody, to form a sandwich antibody–antigen–antibody conjugate upon binding with the capture antibody immobilized at the test line in the presence of luminol, whereupon the nanozyme tagged to the conjugate catalyzes the chemiluminescence conversion of 5-Amino-2,3-dihydrophthalazine-1,4-dione or luminol. In this strategy, the chemiluminescence is captured by camera or cell phone camera which can process the signal intensity using an appropriate mobile application for elegant visual display and graphic analysis. The arrangement and working principle of the strategy is illustrated in Because the impact of the novel nanozyme based innovations on the detection and identification of disease-causing pathogens is shorter detection time, lower detection limits, cost-effective detection, concentration dependent results, and quality devices, point-of-care usage will be enhanced. These improvements in detection and identification of disease-causing pathogens at the point-of-care will benefit accurate diagnosis and responsive therapeutic strategies. Kumawat et al., (2021) predict that nanozymes will play a leading role in the fight against the current and future pandemics . Along wWhile the detection and identification of pathogens has evolved from the historic culture-and-grow methods, through the laboratory-based manual analytical procedures, the rise in pathogen sponsored disease burden, suffering, and deaths over the evolution period, has escalated the need for fast and cost-effective point-of-care methods, giving rise to many paper-based and microfluidic methods of detection and identification of pathogens. Significantly during this period, the PCR and immunoassay methods have contributed many significant gains in some of the most important bio-analytical methodology and strategy characteristics, including simplicity, sensitivity, speed of response, cost, reliability, specificity, and durability. Because many of these methods involved elaborate use of natural enzymes, which are expensive and may suffer from gradual degradation, significantly compromising their shelf life and reliability, the advent of nanozymes, which mimic the catalytic activities of natural enzymes, has brought many improvements in these methodological characteristics. However, nanozymes still lack specificity for pathogens and for these reasons they have been used in combination with methods such as PCR, enzyme-linked immunosorbent assays, paper strips, and microfluidic systems which are pathogen specific.This review presents a synopsis of the details of some of the key methods that have been developed and tested for pathogen detection and identification. To put these developments in perspective, a discussion of the types, classes, structures of pathogens, and the implicated disease burden led to an analysis of the historic and current methods used for pathogen detection and identification. In these discussions, it was necessary to provide some detail to rationalize the developmental trajectories based on the difficulties and challenges of traditional approaches along the six most desirable methodological indicators that are proposed as key in this review. While the culture-and-grow method still predominates in resource constrained environments due to lack of infrastructural and knowledge resources, an increasing number of paper-based methods are coming in to ease the burden. For example, it is still considered to be quite normal and robust that sampling in resource constrained environments is immediately followed by sending samples away for analysis and the waiting periods are major drivers of the disease burden and mortality. This is the area in which paper strips are providing the most needed relief ,137.Because the application of nanozymology is the central theme of focus, the review went into detailed analysis of applications of nanozymes in viral, bacterial, and fungal pathogens, while indicating some of the applications in cancer cell and parasite detection. In this regard, examples of how nanozymes have been used in colorimetric and photothermal colorimetric nanozyme-linked immunoassay detection and identification of bacteria were discussed. Other examples discussed in some detail include a colorimetric microfluidic and a fluorescence microfluidic biosensor, an iron oxide nanozyme colorimetric immunoassay paper strip, and a nanozyme-linked immunoassay chemiluminescence paper strip. The two paper strips were discussed with the background of an innovative different flow rate multiple channel amplified colorimetric nanogold paper strip. The immunoassay paper strip and microfluidic biosensor methodologies are increasingly utilizing the capabilities of real time smart phone data capture and display.Due to their importance emerging from the discussions, a comparative analysis of the various methodologies for detection and identification of pathogens considered in the evolutionary profile presented in this review was conducted along the set of six desirable characteristics—complexity, sensitivity, speed of response, cost, reliability, specificity, and durability. The methodological classes considered in this review include culture-and-grow, manual laboratory, and biosensors. Manual laboratory methods include the enzyme-linked immunosorbent assays and PCR. Biosensors include lateral flow paper strips and microfluidic devices. Whereas Li et al., (2000) recognized target recognition molecules, signal transduction, and the substrate material as the three key functional components of biosensors, the nanozymology biosensors considered in this review focused on pathogens directly and molecular antigens indirectly, the colorimetric, photothermal, chemiluminescence, photoluminescence, surface plasmonic, and fluorescence signal transduction, based on nanozymes mimicking enzymes .It is clear that nanozymology has taken the detection and identification of pathogens to greater heights in terms of the six characteristics identified in this review, including specificity.


To investigate effects of all-inclusive and hierarchical rehabilitation nursing model combined with acupuncture on limb function and quality of life in elderly patients with cerebral infarction during convalescence.  Eighty elderly patients with cerebral infarction who were treated in our hospital (February 2018–February 2020) and met the inclusion and exclusion criteria were chosen as the research objects, and their materials were analyzed in the way of retrospective study. They were equably randomized into observation group and reference group. Based on the acupuncture treatment, the patients in the observation group and the reference group were given routine rehabilitation nursing and all-inclusive and hierarchical rehabilitation nursing respectively for three months. The simplified Fugl-Meyer Assessment (FMA) scores, Modified Edinburgh-Scandinavian Stroke Scale (MESSS) scores, Activities of Daily Living Scale (ADL) scores, and Stroke-Specific Quality of Life Scale (SS-QOL) scores in the two groups before and after intervention were recorded, and the changes of limb function, neurological function, and living quality of the patients in the two groups were analyzed. P < 0.001), markedly lower MESSS score (P < 0.001), and signally higher SS-QOL scores (P < 0.05). After intervention, the observation group achieved obviously higher ADL score than the reference group .  Compared with the reference group, the observation group after intervention achieved prominently higher FMA score ( The all-inclusive and hierarchical rehabilitation nursing model combined with acupuncture can accelerate the recovery of neurological function of the elderly patients with cerebral infarction, enhance the rehabilitation of their limb function, and markedly improve their quality of life. Therefore, this model has referential significance in clinic. Lancet further confirmed that the cerebral stroke was the primary cause of death in China from 1990 to 2017 . ThAll the patients were given routine drug treatment in neurological department and consciousness-restoring resuscitation (Xingnao Kaiqiao) acupuncture. The patients were given acupuncture at the acupoints of Renzhong, Sanyinjiao, Neiguan, Jiquan, Chize, and Weizhong. The needles were inserted with diagonal stabbing and reducing method at the acupoint of Renzhong, with diagonal stabbing and reinforcing method at the acupoint of Sanyinjiao, with straight needling and reducing method at the acupoints of Neiguan, Jiquan, Chize, and Weizhong. The needles were retained at each acupoint for 30 min after the arrival of qi, 1 time/d, and 6 d/week. Six days were a course of treatment, and the patients took a day off before receiving the next course. The acupuncture treatment lasted for 3 months.The reference group was given routine rehabilitation nursing on the basis of acupuncture treatment. The nursing staff assist the patients with rehabilitation training by giving them drug use and diet guidance, movement guidance , and posture nursing.The observation group received all-inclusive and hierarchical rehabilitation nursing on the basis of acupuncture, and the specific steps were as follows. (1) Establishing the rehabilitation nursing teams. The team members were divided into 3 grades, including responsible team leaders, primary nurses, and assistant nurses. The responsible team leaders were the charge nurses or vice-director nurses with more than 10 years of nursing experience, the primary nurses were the nurses with 5–10 years of nursing experience, and the assistant nurses were the nurses with less than 5 years of nursing experience. All team members reviewed related literature on hierarchical nursing and learned from advanced experience in the manner of evidence-based nursing. In addition, the responsibility of each grade was specified according to the characteristics of the patients with cerebral infarction in our hospital, so as to clarify the individual responsibility. (2) Establishing the shift system. The nurses of different grades were divided into 3 nursing groups according to their educational level, specialized level, communication ability, and other factors. The 8-hour continuous shift system was implemented, and the team leaders needed to communicate with the nurses and coordinate the scheduling to ensure the integrity and continuity of the rehabilitation nursing. (3) Training. The training contents, mainly including relevant systems, nursing knowledge and professional skills about the rehabilitation nursing of the patients with cerebral infarction, were arranged in accordance with the rehabilitation nursing requirements of each grade, so as to improve the nursing staff's ability of individualized analysis and judgment of the patients' clinical symptoms, monitoring parameters, and nursing key points. (4) Division of work. ① Assistant nurses were mainly responsible for the patients' basic nursing, such as morning and evening nursing, ward ventilation, and bedding changing, and they should improve the quality of basic nursing to provide the patients with good conditions of recovery. ② Primary nurses were mainly responsible for the patients' routine nursing, such as injection and puncture, and they should improve the eligible rate of the routine nursing operation to ensure the effective progression of the routine nursing. ③ The charge nurses were in charge of the patients with cerebral infarction in all respects, including the health education, mental nursing, recovery guidance, and management of daily life. The elderly patients with cerebral infarction were more likely to develop such adverse emotions as anxiety and irritability due to the disease factors, which affected their compliance with the rehabilitation nursing. Therefore, the charge nurses need to carefully observe the patients' emotional changes and solve their psychological problems in time to ensure that the patients kept good psychological states and received the rehabilitation nursing with positive and healthy attitudes. The charge nurses provided guidance on the patients' simple activities in their daily lives and rehabilitation training in standing or walking according to the patients' conditions, regularly checked the patients' physical conditions, and formulated targeted nutritious diets to strengthen their body immunity. The elderly patients with cerebral infarction stayed in bed for a long time, so they had difficulty in turning over and were likely to be complicated with dermatosis, such as pressure ulcer. Hence, the charge nurses needed to offer the patients posture nursing and tried to keep them in lying position on the health side. When the patients took lying position on the affected side, the charge nurses should help the patients to turn over to avoid pressure ulcer and other complications. After the patients being admitted, their affected limbs were wiped with warm water every day, and they were given the massage with 50% alcohol before going to bed at night to prevent their joints from deforming and being stiff. ④ The vice-director nurses were mainly responsible for the patients' rehabilitation training and difficult nursing operations. After the patients' signs stabilized, the reasonable, scientific, and efficient rehabilitation plans were formulated on the basis of their conditions, and the patients were informed of relevant matters needing attention and methods of rehabilitation training to enhance their cooperation. Then, the rehabilitation training was conducted, and the patients were instructed to move limbs while lying in bed, like stretching out the hands and bending the elbows. After the patients could get out of bed, they did standing balance exercise, torsion training, Tai Chi, and other daily training with the help of vice-director nurses. The nurses should control the intensity of activities, follow the principle of gradual improvement, and timely adjust rehabilitation plans according to the patients' recovery. The vice-director nurses should encourage the patients to read books, listen to music, and watch TV, because proper sensory stimulation was conducive to improving the patients' language sensibility and speeding up the recovery of neurological function. The vice-director nurses could massage the patients' upper limbs with kneading method and lower limbs with rolling method. The massage strength should be from light to stronger and the patients' reaction should be observed.Fugl-Meyer Assessment (FMA)  was usedModified Edinburgh-Scandinavian Stroke Scale (MESSS)  was usedActivities of Daily Living Scale (ADL)  were adoThe Stroke-Specific Quality of Life Scale (SS-QOL)  was used2 test and t test. When P < 0.05, the differences were considered statistically significant.This study adopted SPSS20.0 as the data processing software and GraphPad Prism 7  as the graph drawing software. The study included count data and measurement data and used XP < 0.001; Compared with the reference group, the observation group achieved prominently higher FMA score after intervention (P > 0.05). Compared with the reference group, the observation group had prominently higher FMA score of upper limbs after intervention .No prominent difference in the FMA score of upper limbs between the two groups before intervention was found . Compared with the reference group, the observation group had markedly higher FMA score of lower limbs after intervention .No obvious difference in the FMA score of lower limbs between the two groups before intervention was found . Compared with the reference group, the observation group had much lower MESSS score after intervention .No prominent difference in the MESSS score between the two groups before intervention was found . Compared with the reference group, the observation group had markedly higher ADL score after intervention .No remarkable difference in the ADL score between the two groups before intervention was found ; after intervention, the observation group achieved obviously higher ADL score than the reference group ; the patients in the observation group had higher living wills and better quality of life. In the long term, the all-inclusive and hierarchical rehabilitation nursing model can effectively reduce the incidence of nurse-patient disputes. This study provides more reference for reducing the medical burden of cerebral infarction in China.Compared with the reference group, the patients in the observation group had better rehabilitation efficacy in neurological and limb function, because the observation group received efficient nursing intervention on the basis of acupuncture. At this stage, the nursing intervention commonly used in clinics during convalescence mainly aims at the patients' limb rehabilitation, relieving their clinical symptoms and recovering their living ability. Although such nursing intervention has certain effects, it ignores the patients' subjective initiative. Indeed, it does not take the patients with cerebral infarction as the center of rehabilitation nursing . AlthougAt present, the mainstream rehabilitation nursing still has some deficiencies, impeding tit from exerting maximum efficacy, so it is urgently needed to be improved. The all-inclusive and hierarchical rehabilitation nursing model combined with acupuncture can accelerate the recovery of neurological function of the elderly patients with cerebral infarction, enhance the rehabilitation of their limb function, and markedly improve their quality of life. Therefore, this model has referential significance in clinics.


The present review represents the outstanding and promising recent literature reports (2017–2022) on nanoparticle-based formulations developed for anticancer therapy with photodynamic therapy (PDT), photosensitizers, and chemotherapeutics. Besides brief descriptions of chemotherapeutics’ classification and of PDT mechanisms and limitations, several examples of nanosystems endowed with different responsiveness  and peculiarity  are described, and for each drug combination, in vitro and in vivo results on preclinical cancer models are reported.The widespread diffusion of photodynamic therapy (PDT) as a clinical treatment for solid tumors is mainly limited by the patient’s adverse reaction (skin photosensivity), insufficient light penetration in deeply seated neoplastic lesions, unfavorable photosensitizers (PSs) biodistribution, and photokilling efficiency due to PS aggregation in biological environments. Despite this, recent preclinical studies reported on successful combinatorial regimes of PSs with chemotherapeutics obtained through the drugs encapsulation in multifunctional nanometric delivery systems. The aim of the present review deals with the punctual description of several nanosystems designed not only with the objective of co-transporting a PS and a chemodrug for combination therapy, but also with the goal of improving the therapeutic efficacy by facing the main critical issues of both therapies . Therefore, particular attention is paid to the description of bio-responsive drugs and nanoparticles (NPs), targeted nanosystems, biomimetic approaches, and upconverting NPs, including analyzing the therapeutic efficacy of the proposed photo-chemotherapeutic regimens in in vitro and in vivo cancer models.  S. S113]. As regards our literature survey on NP-mediated TOPII inhibitors delivery in combination with PSs, no examples of compounds other than DOX were judged notable for the scope of the present review.® or Doxil® for the liposomal formulation of the PS verteporfin and of DOX, respectively), while no human studies on combination of PDT and chemotherapy are reported. Therefore, an open question remains on the effective limitations that prevent the translation of the proposed approaches from the laboratory scale to clinical trials.In the last two decades, nanotechnological approaches and continuous research on biocompatible materials for drug delivery purposes have completely changed the perspectives of anticancer drug administration. Even if majorly at the pre-clinical stage, the use of precisely designed nanosystems has revealed their huge potential not only in increasing drug bioavailability, but also in reducing side effects and allowing a controlled drug release in the TME, thus counteracting important criticisms of cancer therapy. As reviewed in the present paper, even in the context of chemotherapy and PDT, difficulties in the administration of hydrophobic compounds as well as undesired side effects have been attenuated by drugs inclusion in nanometric delivery systems, which allowed the simultaneous co-transport of chemotherapeutics and PSs to improve tumor delivery, drugs timing, and consequently efficacy. Overall, the examples presented in the review highlighted how an accurate selection of materials endowed with bio-responsiveness or selected for possessing unique prerogatives, sometimes combined with a smart drug design  or biomimetic approaches, could effectively maximize tumor reduction in in vivo cancer models with respect to single treatments or with respect to free drug combinations administered in the absence of delivery systems. Nonetheless, notwithstanding the impressive numbers of reports in the field, especially in the last five years, reported in vitro and in vivo preclinical results from completed or ongoing clinical trials foreseeing the use of nanoparticle-based drug formulations still remain restricted to already approved products (e.g., Visudyne


We are delighted to introduce a very special Research Topic of Frontiers in Psychology. Eating disorders (ED) and weight disorders (WD) are severe illnesses in which especially, but not exclusively, young people go through critical disturbances in their eating behaviors and related thoughts and emotions. Given the severity and scope of obesity and unhealthy diets on health, these disorders could have serious outcomes on health, quality of life, and global economy if prevention mechanisms are not arranged. Despite the research conducted up to date, ED and WD are a subject of permanent relevance due their life-threatening condition. In this regard, the field is matter of continuous update aiming to increase the knowledge, improve treatment options, incorporate new research methods and technology, reaching understudied populations, and understanding the impact of external factors. This Research Topics therefore aims to be a contribution to the field of ED and WD; moreover, given the situation of the recent pandemic of COVID-19, which its outbreak on depression, anxiety, distress, insomnia, and post-traumatic stress have been explored . There is also a review article  and a study protocol . The Research Topic also includes a correction  of the corresponding study.This Research Topic includes 16 articles, 13 of them are based on original research coming from different countries . Most of the original research are quantitative studies, and only one study is qualitative  leads to ED symptomatology both directly and indirectly through a sequential mediating path. Specifically, high levels of EI were associated to greater self-esteem, which subsequently reduces anxiety, thus predicting lower levels of ED symptomatology. Marco et al. estimated the mediating role of meaning in life in the relationship between emotional dysregulation and ED pathology in three Spanish samples with diverse risk . They found that meaning in life acted as a mediator between emotional dysregulation and the main symptoms of ED. These results highlight the relevance of considering meaning in life as a variable in the onset and maintenance of ED. Fan et al. proposed a model to explain the relationship between diverse types of weight stigma and quality of life among children in Hong Kong. They found associations between the weight stigmas and both child-rated and parent-rated quality of life. Also, weight stigma and weight-related self-stigma mediated the association between body weight and children's quality of life. Finally, Hampshire et al. describe in their systematic review, that parenting styles characterized by high levels of control and low levels of responsiveness were associated to disordered eating among young adults such as unhealthy weight control behaviors. The association was frequently indirect and mediated by several underlying mechanisms including emotional reactivity, lowered self-competence and offspring psychological distress. These results suggest that parenting styles, parents' difficulties in managing their emotions, their perceived competence, and their psychological distress influence the development of disorders eating of their young-adult children.Four papers have explored the potential role of psychological resources  and psychosocial stressors  on the development and maintenance of these disorders. open science  to Spanish caregivers and adolescent patients with ED. The intervention is based on emotional regulation and eating behaviors, with an emphasis on behavior change strategies. The objective is to generate cognitive and behavioral changes and strengthen relationships with family and friends to help the transit from inpatient care to daily life and to prevent relapses.Following the paradigm of  UNESCO, , Quiles Sierra et al. explores the emotional processing of food images comparing three clinical groups and a normal-weight comparison group. ED patients are characterized by more negative appraisal, less emotional dominance, and a higher level of arousal when processing food-relevant information. Negative affect mediated the relationship between eating symptomatology and emotional processing. This supports the transdiagnostic model of ED, and the use of similar treatments for different conditions.The study by Sepúlveda et al. identified some specific correlates associated with the development of an eating disorder. The model showed that patients' drive of thinness and self-oriented perfectionism, together with the decrease of triiodothyronine (T3) hormone and higher levels of anxiety increase adolescents' risk of developing an ED. Both the increase in fathers' emotional overinvolvement, and the decrease in fathers' depression and mothers' anxiety were specifically associated to the onset of an ED.To generate an exploratory bio-psychological-familial model, the case-control study conducted by Obeid et al. explored a complex model of shared pathways integrating various theoretical facets for eating and weight disorders in a large community-based sample of male and female adolescents. Higher levels of stressors , dysregulated eating and poor body image all directly influenced higher weight status. It confirmed the concept that socio-environmental factors would be associated with body image and dysregulated eating, mediated by the complex interplay of psychological and behavioral factors.Using structural equation modeling, El Archi et al. explored the psychological factors associated with binge eating and food addiction, and the role of emotion dysregulation, alexithymia and personality dimensions in the association between attention-deficit/hyperactivity disorder and addictive-like eating. Emotion dysregulation, conscientiousness, agreeableness, and neuroticism are total mediators of this association, with alexithymia being a partial mediator. Negative emotion leads to food intake, increasing positive emotion seeking especially if associated with emotion dysregulation, leading to the development of an addiction. Despite the controversies of the concept of food addiction, authors suggested that certain foods may lead to addictive-like eating that could contribute to overeating and obesity.Among bariatric surgery patients, Ziser et al., which explored the barriers to behavior change in a group of 16 parents of preschool children with overweight and/or obesity. They were identified through a primarily inductive content analysis conducted through an interview. The underestimation of health consequences of overweight and obesity, and a deficient awareness on the problem were the main barriers. The use of the motivational interview in the pediatric environment are suggested to address these topics.The influence of the familial context was investigated in the qualitative study of Lizana-Calderón et al. analyzed the psychometric properties of the Eating Disorder Inventory-3 . Based on the diet-related lifestyle change during the pandemic, among women with symptoms of ON, findings showed an aggravation of disordered eating thoughts and behaviors and suggested that social media could be contributing factor to this exacerbation. Consistently, the study of Corno et al. described a tendency to report a higher frequency of disordered eating behaviors during the COVID-19 pandemic. Weight concerns predicted an overall increase in the frequency of restrictive behaviors, and body dissatisfaction was found to be associated with a self-perceived increase of emotional eating frequency among adult women who were never diagnosed with an ED. Altogether, both studies suggest that lifestyle change due to the COVID-19 pandemic influenced ED and WE symptoms.Finally, two papers aimed to assess the impact of the COVID-19 pandemic on ED and WD symptoms. A variety of populations from Asia, Europe, North America, and South America including clinical and community-based samples are represented across the studies. Diverse research designs were considered to assess mediational mechanisms underlying ED and WD symptoms, as well as emotional processing of ED-related stimuli, and specific correlates associated with the onset of ED among clinical samples. Additionally, factors associated with binge eating and food addition have been explored in bariatric surgery patients, and qualitative data describes barriers to behavior change among parents of overweight/obese children. This Research Topic also includes the validation of three measures across Hispanic samples, and a study protocol of an intervention aimed to improve the transit of inpatient care to daily life. Finally, two studies described the impact of the COVID-19 pandemic on disordered eating behaviors. We expect that this Research Topic represents a contribution to the ongoing scientific knowledge about ED and WD.All authors listed have made a substantial, direct, and intellectual contribution to the work and approved it for publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Anemia is one of the most frequent diseases worldwide, affecting one-third of the general population. Anemia in general and in particular, iron-deficient anemia (IDA), has been associated to a higher risk of thrombotic manifestations, including ischemic stroke and cerebral venous thrombosis (CVT), as well as systemic extra-cerebral arterial and venous thrombosis. Despite these data, anemia is seldom considered as an etiological factor of stroke. An individual case encompassing all known neurovascular and systemic arterial and venous thrombotic manifestations related to IDA is presented with the focus on clinical reasoning issues in the diagnostic pathways, starting from the neuroradiological signs. The main questions have been identified and addressed in a narrative review of the most relevant data in the literature from a pragmatic and clinical viewpoint. The presented case concerns a 46-year-old man admitted to the Stroke Unit because of acute ischemic stroke with multiple thrombi in large intracranial and extracranial vessels, multifocal ischemic lesions in several arterial territories and the concurrent finding of asymptomatic CVT, pulmonary embolism with lung infarction and aortic thrombosis. An extended diagnostic work-up excluded the main etiologies , except for a severe IDA, such as to require blood transfusions followed by anticoagulant therapy for the several thrombotic manifestations. Neuroimaging and systemic vascular findings have been analyzed, and the main issues proposed by the case in the diagnostic pathway have been identified and discussed in a pragmatic clinical road map reviewing the data provided by the literature. Conclusions: IDA is a common but treatable condition that, independently or synergically, may increase the risk of thrombotic events. The diagnostic and therapeutic approach has not yet been defined, and each case should be individually addressed in a pragmatic clinical road map. Acute ischemic stroke with neuroimaging patterns suggesting an embolic source is a not rare occurrence in both young and old patients. Usually, the underlying embolic mechanism is inferred because of the presence of multiple ischemic lesions in more than one arterial vascular territory, but it is very rare to find a thrombus in a large vessel like a “smoking gun” in the diagnostic pathway looking for an etiology of the stroke. It is even rarer to find the simultaneous presence of thrombi in multiple arteries and the association of arterial and venous thrombosis as well as cerebral and extra-cerebral thrombosis. The current etiological classifications of ischemic stroke do not include such cases in any category, and the diagnostic pathway is not clearly defined, shared and codified. Similarly, the most appropriate therapeutic approach to these situations has not been defined, and each case has an individual management. Among the different possible etiologies are acquired or hereditary prothrombotic conditions as well as common diseases that have a rare presentation in which stroke is the first manifestation; not infrequently, the diagnostic work-up does not lead to the identification of a certain proof, allowing to collect only a series of faint clues.A 46-year-old man was referred to the Emergency Department (ED) in the afternoon because of unsteadiness, vertigo and nausea, reporting the awareness of the symptoms at awakening, around ten hours before the consultation; his last time well was almost 24 h before. His past medical history was notable for two episodes of head concussion without clinical consequences and brain lesions on CT imaging, the recent finding of increased value of glycated hemoglobin and a long-lasting IDA, erratically treated with iron oral supplementation. The first blood laboratory assessment in ED confirmed the microcytic anemia without other relevant findings: hemoglobin 8.3 g/dL , RBCs 4.26 × 106/µL , hematocrit 28.6% , MCV 67.3 fL , MCH 19.5 pg , MCHC 28.9 g/dL , platelets 211 × 1000 µL , fibrinogen 188 mg/dL , D-dimer 2485 ng/mL , regular liver and kidney functions, absence of electrolytic disturbances and normal inflammatory markers. The initial neurological examination was remarkable for impaired alertness , dysarthria and left limbs ataxia with a global National Institute of Health Stroke Scale (NIHSS) score of 11. The patient underwent an unenhanced brain CT scan, whose findings were significant for multiple subcortical hypodensities in both cerebellar hemispheres and vermis, which was highly suggestive of subacute ischemic lesions in the territory of bilateral posterior inferior cerebellar artery (PICA) and left superior cerebellar artery (SCA) .A rounded hypodense lesion partially adherent to the aortic arch wall and partially floating within the lumen of the aortic arch, in the segment between the origin of the left common carotid artery (CCA) and the left subclavian artery (SA);A similar smaller hypodense rounded structure partially adherent to the posterior wall of the brachiocephalic artery (BCA);A progressive reduction in intensity of contrast filling of the right vertebral artery (VA) starting from the V2–V3 transition, leading to a complete occlusion in the V4 segment;A lack of contrast filling of the right lateral dural venous system involving transverse sinus (TS) and sigmoid sinus (SS) and also reaching the jugular bulb and the proximal extracranial segment of the right internal jugular vein (IJV).Therefore, an arch-to-vertex CT-angiography (CTA) was performed, and it revealed the following:The intraluminal arterial hypodensities described in   are suggThe presence of multiple subacute ischemic lesions already evident in the brain CT at admission excluded intravenous thrombolysis and endovascular treatment. Therefore, the patient was admitted to the Stroke Unit for acute stroke care and further investigations and started single antiplatelet treatment and low-molecular weighted heparin at a prophylactic dose. At the admission, the patient shown a stepwise neurological worsening, and the final clinical examination at 12 h from the admission was remarkable for a right facial–brachial hemiplegia, cerebellar dysarthria, dysphagia, inconstant limitation on left lateral gaze without nystagmus and a left cerebellar syndrome. A brain MRI was performed at about 15 h from the admission, confirming the presence of multiple subacute ischemic areas, located in the left parasagittal vermis and in the medial–superior region of both cerebellar hemispheres, and outlining other subacute infarctions involving the right frontal and parietal lobe  and the left paramedian medullary region . MoreoveFurther tests were carried out trying to identify other prothrombotic factors and to clarify the etiology of the clinical picture. The extended laboratory investigations included normal total (360.6) and unsaturated iron-binding capacity (283.6), transferrin saturation (21.3%), serum ferritin , serum iron  and transferrin (284 mg/dL). The fecal occult blood test was negative; nocturnal paroxysmal hemoglobinuria and hemolytic anemia were excluded, and the analysis of hemoglobin subgroups resulted within a normal profile. Antibodies anti-double stranded DNA, anti-mitochondrial (AMA), anti-smooth muscle (ASMA), anti-nuclear (ANA), anti-phospholipid, anti-deamidated gliadin peptide and anti-transglutaminase were within the normal range; the main onco-markers  did not express any significant pathological increase. Anti-thrombin levels were within the normal range, the search for a mutation of Factor V-R506Q was negative; a heterozygote genotype was identified for MTHFR-C677T, and an insignificant increase in homocysteine values was found. Transthoracic and transesophageal echocardiography as well as 24 h ECG monitoring were unremarkable. Conversely, a thoracic-abdominal CT scan highlighted thromboembolisms involving also the sub-segmental branches of both pulmonary inferior lobes and an 8 mm free thrombus adherent to a crescent-shaped atheroma of aorto-iliac junction and floating within the lumen .At 30 days from the symptoms’ onset, the neurological condition was substantially unchanged. A follow-up brain MRI highlighted a regular evolution of the ischemic lesions and confirmed the persistent occlusion of the right V4 VA. Moreover, the brain MRI showed as unchanged the white matter hyperintensities in the external capsula and centrum semiovale  togetherConversely, the right TS, SS and IJV CVT was recanalized in the follow-up MRA , suggestFinally, a whole-body G18-FDG PET did not show abnormal metabolic activities except for both basal pulmonary areas and for the gastro-duodenal region. The former one was explained by the pleural effusion secondary to pulmonary embolism, while the latter was explored with endoscopic and histologic studies that did not reveal abnormalities.During the hospital stay, the neurological condition was substantially unchanged. The severity of anemia in the acute phase requested a blood transfusion (two units of packed red blood cells), which was followed by iron intravenous infusion and vitamin replacement. The hemoglobin level progressively increased until a persistent stabilization above 12 g/dL. LMWH dose was increased to an anticoagulant posology after blood transfusion and insulin therapy was set up.Even though a large panel of investigations was performed, we were not able to identify other genetic or acquired prothrombotic factors but IDA nor a cause of bleeding that could explain the microcytic anemia. In consideration of the multi-district thrombosis, involving also the pulmonary district, after the hospital discharge, oral anticoagulation with rivaroxaban 20 mg was started for venous thromboembolism (VTE) treatment. A 12-month follow-up did not raise any other etiology.This clinical case raises several questions both in the diagnostic steps and in the therapeutic choices and has almost unique characteristics, joining together in the same patient simultaneous arterial and venous thromboses in multiple sites. Starting from the symptoms, which led the patient to the ED, the first neuroimaging studies in ED made already evident the clinical complexity related to the presence of multiple sites of thrombotic occlusions in the large arteries supplying the brain and in the cerebral venous sinuses. While arterial thrombosis was symptomatic and its acute timing was easily defined by the presence of a recent multifocal ischemic stroke, CVT was an incidental and asymptomatic finding and, as such, undated. Only later, the resolution of CVT with anticoagulation did support its recent timing. At this step, even with a history of chronic IDA, the causal hypotheses for simultaneous arterial and venous thrombosis were substantially limited to genetic or acquired prothrombotic conditions, which is in accordance with the existing stroke classifications. A genetic thrombophilia would have had a lower probability because of the simultaneity of thrombotic findings and the absence of a previous history of thrombotic events, even minor ones. Moreover, the simultaneous involvement of the venous and arterial sides virtually excluded major cardioembolic sources as atrial fibrillation (AF), and a paradoxical embolism mechanism would not be able to take account of the etiology of venous thrombi. However, the patient did not have a defect of the interatrial septum, and two weeks of heart monitoring did not identify atrial fibrillation.As a general construct, the presence of multiple cerebral ischemic lesions with an embolic pattern and without an identifiable known etiology leads to the consideration of the embolic stroke of undetermined source (ESUS)  concept.In the diagnostic work-up of cryptogenic embolism, the co-occurrence of venous and arterial thrombi in cerebral circulation was suggestive for two main diagnostic hypotheses, i.e., antiphospholipid antibodies syndrome (APS) and cancer-associated stroke (CAS). APS is a disease classically associated to many thrombotic manifestations in arterial and venous districts ,5,6, butCAS was a meaningful diagnostic hypothesis, and several issues may raise it in the context of the diagnostic work-up of cryptogenic embolism . Indeed,Moreover, a multiple scattered lesion pattern, often reported in CAS, was associated to higher D-dimer values ,18,19. TUnfortunately, these clues have some limitations. In particular, because stroke itself is frequently associated to an increased D-dimer, its value alone is not sufficient for this purpose. D-dimer is a more general marker of an activated coagulation system, and the presence of thrombi and ischemic lesions in the presented case is usually associated to the finding of increased D-dimer values independently from the cause of the thrombosis. Another surrogate marker of the pro-thrombotic role of cancer is the detection of High-Intensity Transient Signals (HITS), also known as microembolic signals, on Transcranial Doppler (TCD) , but in The finding of multiple thrombi in large vessels, both arteries and veins, in the presented case deserves a dedicated consideration as a potential clue for a hidden malignancy, since hypercoagulability is the most relevant mechanism of CAS  and knowIf the diagnostic pathway of the presented case was analyzed according to the advanced diagnostic protocol  proposedIndeed, the neuroimaging pattern of the patient does not fulfill a single category, but it combines features belonging to the three DWI patterns and leaves open the possibility of multiple etiologies except for branch occlusive disease. The multiplicity of lesions  and the recent and past timing of the findings is hard to lead back to a single category. In the presented case, a hidden malignancy was extensively looked for, but all studies were negative in this regard both in the acute phase and in the 12-month follow-up. Other, even rare, recognized causes of stroke and thrombosis have been investigated and excluded. At this point, IDA was considered with more attention, focusing on the described mechanisms linking IDA and thrombosis and on the literature data.p < 0.001) and the odds ratio (OR) of previous IDA for subjects with a VTE was 1.43 : 1.10–1.87) compared with the controls. The mechanisms of the association between IDA and increased thrombotic risk, both arterial and venous, however, are not fully clarified, and the simultaneous finding of multiple arterial and venous thrombotic events in the same individual has been reported only in an anecdotal way and often evokes etiologies different from anemia in the differential diagnostic pathway. Moreover, the prothrombotic role of IDA therefore appears to be considerably underestimated in neurovascular clinical practice, and the diagnostic and therapeutic pathways of these patients have yet to be defined, raising still unanswered questions.Anemia is one of the most frequent diseases worldwide and, although the great differences due to age, sex and geographical distribution make it difficult to assess the real incidence, it roughly affects one-third of the general population ,26,27. AThe association between IDA and thrombotic risk is further increased by thrombocytosis. To this regard, the development of an elevated platelets count, probably due to the hyperactivity of erythropoietin in anemic state, has been considered one of the potential thrombotic mechanisms . Indeed,A further consideration relating to this and other attempts at the phenotyping of cryptogenic embolism, both before and after the introduction of the ESUS concept, is that none of these includes the finding of thrombi in large vessels as a distinctive element nor the coexistence of arterial and venous thrombosis or the simultaneous presence of cerebral and systemic thrombosis. In some of these classifications, the cerebral and systemic ischemic lesions are considered, but the identification of thrombi in the vessels is not among the classifiable items. Interestingly, also, the more detailed subtyping proposed by the critics of ESUS concept  includedTo our knowledge, there are no previous case descriptions of simultaneous arterial and venous thrombosis in patients with IDA as isolated prothrombotic factors. However, if the prothrombotic power of anemia has been widely accepted, how and if it may independently be responsible for arterial and/or venous thrombosis has not been completely clarified. Nevertheless, our case confirms how IDA might be considered, de facto, a strong prothrombotic factor, and it is able to produce a multi-district systemic thrombosis in a young patient without other relevant medical conditions. Thus, it should be important to point attention to a treatable disease, such as IDA, and its strict association with a life-threatening condition, such as cerebral and systemic thrombosis. Moreover, if we consider that vascular events may be related to hardly modifiable risk factors, anemia falls into the category of treatable conditions, and its correction may affect the outcome of many patients. A key issue might be that despite or due to its high incidence in the general population, anemia could be underdiagnosed. Furthermore, a poor compliance to long-term iron replacement could influence the success of the therapy, as happened to our patient. Surely, raising the awareness on the importance of an earlier identification of anemia and its causes might be a critical target, although it was not been demonstrated that treating anemia reduces the risk of thrombotic events.This unique clinical case raised attention regarding a common but treatable condition that, independently or synergically, may increase the risk of thrombotic events in all body districts and across all ages. IDA should be considered as a cause of ischemic stroke, mainly if it is associated with the evidence of thrombotic occlusion of large vessels and with systemic and venous thrombosis.