As global interest in the therapeutic potential of cannabis and its’ derivatives for the management of selected diseases increases, it is increasingly imperative that the toxic profile of cannabinoids be thoroughly understood in order to correctly assess the balance between the therapeutic risks and benefits. Modern studies across a number of jurisdictions, including Canada, Australia, the US and Europe have confirmed that some of the most worrying and severe historical reports of both congenital anomalies and cancer induction following cannabis exposure actually underestimate the multisystem thousand megabase-scale transgenerational genetic damage. These findings from teratogenic and carcinogenic literature are supported by recent data showing the accelerated patterns of chronic disease and the advanced DNA methylation epigenomic clock age in cannabis exposed patients. Together, the increased multisystem carcinogenesis, teratogenesis and accelerated aging point strongly to cannabinoid-related genotoxicity being much more clinically significant than it is widely supposed and, thus, of very considerable public health and multigenerational impact. Recently reported longitudinal epigenome-wide association studies elegantly explain many of these observed effects with considerable methodological sophistication, including multiple pathways for the inhibition of the normal chromosomal segregation and DNA repair, the inhibition of the basic epigenetic machinery for DNA methylation and the demethylation and telomerase acceleration of the epigenomic promoter hypermethylation characterizing aging. For cancer, 810 hits were also noted. The types of malignancy which were observed have all been documented epidemiologically. Detailed epigenomic explications of the brain, heart, face, uronephrological, gastrointestinal and limb development were provided, which amply explained the observed teratological patterns, including the inhibition of the key morphogenic gradients. Hence, these major epigenomic insights constituted a powerful new series of arguments which advanced both our understanding of the downstream sequalae of multisystem multigenerational cannabinoid genotoxicity and also, since mechanisms are key to the causal argument, inveighed strongly in favor of the causal nature of the relationship. In this introductory conceptual overview, we present the various aspects of this novel synthetic paradigmatic framework. Such concepts suggest and, indeed, indicate numerous fields for further investigation and basic science research to advance the exploration of many important issues in biology, clinical medicine and population health. Given this, it is imperative we correctly appraise the risk–benefit ratio for each potential cannabis application, considering the potency, severity of disease, stage of human development and duration of use. Cannabinoid genotoxicity is not controversial and is widely acknowledged by both government drug regulators and the cannabis industry. The official prescribing information for both Sativex (Δ9-tetrahydrocannabinol (THC)—cannabidiol) and Epidiolex (cannabidiol) registered with the US Food and Drug Administration (FDA), the European Medicines Agency (EMA) and the Medicines and Health Care Products Regulatory Agency (MHCPRA) of the UK ,2 carry However, while cannabinoid genotoxicity has been well established experimentally in a variety of different in vitro systems ,17,18,19With rising interest in the pharmaceutical use of cannabis for the management of selected diseases and a strong commercial and popular interest in the therapeutic potential of cannabis derivatives, it becomes increasingly important that the toxic profile of cannabinoids is thoroughly understood in order to parse the classical balance between the therapeutic risks and benefits correctly and astutely. Inherently, this risk–benefit ratio must be correctly appraised for each potential cannabis application, considering the potency, severity of disease, stage of human development and duration of use. The correct and astute appraisal of cannabinoid toxicity becomes even more important in the case of cannabinoids where, at the time of writing, reports from both the Institute of Medicine and the National Academy of Sciences described the objective evidentiary basis underpinning cannabis prescription as often “weak” to “moderate at best”, and most of its hypothetical indications as being of anecdotal level only, and so of weak evidentiary power ,21.Contrariwise, strong epidemiological evidence from many jurisdictions showed the likely involvement of cannabinoids in many cancers, dozens of serious birth defects and cellular and organismal aging ,52,53,54While it is acknowledged that some historical literature assessing the cannabis morbidity associations produced data showing no effect, the field is plagued with numerous methodological difficulties, including the conduct of many widely quoted studies in earlier eras when cannabis was of a much lower THC potency and the increase in the community exposure since that time, in terms of not only the prevalence of cannabis consumption but the intensity of cannabis consumption. In some widely quoted studies , the syssimultaneously, these features can be expected to operate synergistically as a relatively abrupt switch where severe adverse genotoxic and neurotoxic outcomes relatively suddenly become commonplace due to the underlying exponential dose-response effects [Importantly, in attaining a cannabis dose-response threshold, there has been an increase in the cannabis exposure for some communities, arising not only from an increased prevalence in cannabis consumption but from an increased potency and frequency of use ,57, wher effects ,57.simultaneously implies a major paradigmatic shift in the community exposure from a relatively low level to a much higher region where genotoxic outcomes become more common.It is well established that the trend of cannabinoid exposure in both the US and Europe has been upwards and positive (increasing) in recent decades. This rising trend carries two major implications. Firstly, in the US, it has been shown that the prevalence of cannabis use has increased, the rate of daily cannabis use has more than doubled ,62 and tSecondly, it would be expected that the rising trend in cannabinoid exposure shows a simple bivariate statistical correlation with any rising trend in the population with ill health. Accordingly, recent research has established a robust relationship with multisystem tumorigenesis and teratogenesis, which is robust to multivariable adjustment, is verified in combined formal spatiotemporal analyses and satisfies the modern quantitative criteria of causality with very similar results reflected in both the US and European datasets ,45,46,67It must be underscored that the triple convergence of cannabinoid carcinogenesis, cannabinoid teratogenesis and the cannabinoid acceleration of aging together forms strong and theoretically robust evidence for a clinically and highly significant genotoxicity  severe eFurthermore, both in vitro and clinical studies implicate many different cannabinoid moieties, suggesting that genotoxicity is a class effect shared by many cannabinoids ,70—a feaFor the reader who is unfamiliar with this epidemiological literature, it should be pointed out that most of the modern epidemiological studies referred to are not just observational ecological studies of convenience which happen to show a particular association. Many of the best studies used a formal space–time analysis and the quantitative tools of causal inference to introduce a pseudo-randomized quasi-experimental paradigm from which it is entirely appropriate to invoke causal associations ,80,81,82It is the purpose of the present paper to explore the manner in which the diverse recent laboratory and clinical results, which are the outcomes of the three primary expressions of cannabinoid genotoxicity in cancerogenesis, teratogenesis and aging, are explained using cannabinoid epigenotoxicity and the mechanistic insights which follow an improved understanding and appreciation of the magnitude and breadth of the scope of the epigenotoxic profile of cannabinoids. Since this is only beginning to be explored, the present review is necessarily limited to an introductory oversight, but it does lead to the formulation of a broad ranging experimental investigation into the ways in which these insights can be developed and explored further with far reaching implications across the spectrum of clinical medicine and the basic biological sciences.There is impressive overlap between the US and European data for cannabis exposure and tumors. A recent review of 28 US cancer tumors that were significantly associated with Δ9THC included acute myeloid leukemia, breast, oropharynx, thyroid, liver, pancreas, chronic myeloid leukemia, testis and kidney . The canSimilarly, in a review of 40 European cancers, 27 tumors were related to various metrics of cannabis exposure, including daily use. The tumor overlap included all cancers (excluding non-melanoma skin cancer), oropharynx, the four major leukemias and Hodgkin’s and non-Hodgkin’s lymphoma, liver, pancreas, brain medulloblastoma, anus, kidney, thyroid, testis (seminoma and non-seminoma), ovary and ovarian germ cell tumors. They also identified hepatocellular, skin melanoma, mesothelioma, Kaposi sarcoma, penis, prostate, vulva and vaginal cancers .Hence, there is impressive overlap between the US and European data for those tumors which are listed as common.It is mechanistically noteworthy that, for several of the tumors aforementioned, chromosomal translocation is an important and well-established pathway in their oncogenesis. The presumed mode of action here is to constitutively activate the proto-oncogenes or suppress the tumor suppressor genes. These comments apply particularly to acute myeloid and lymphoid leukemias (AML and ALL) and to testicular cancer ,85,86,87When 62 congenital anomalies were tracked longitudinally across the US, 45 were shown to be related to the metrics of cannabis exposure, particularly those from the cardiovascular, chromosomal, gastrointestinal, limb, urinary, body wall and face .When a series of 95 European congenital anomalies were studied, 89 were shown to be relatable to the various metrics of cannabis exposure and were related particularly to the anomalies affecting the cardiovascular, gastrointestinal, uronephrological and central nervous systems, as well as the chromosomal, limb, body wall, face and general  .Again, a significant overlap was shown where the anomalies listed were in common .It is of interest that if one adds the lengths of the chromosomes directly impacted by these chromosomal anomalies together, it results in 388 of the 3000 megabases of the human genome, or 12.9%.The data from the twelve separate empirical streams both independently and more strongly collectively provide a convincing empirical case for cannabis-induced accelerated aging. These include hepatotoxic , immunolA recent detailed report from a large electronic health record database in Hawaii showed that cannabis users were subject to elevated rates of an impressive array of acute illnesses, including myocardial infarction, stroke, acute bronchitis, cyclic vomiting, injuries, poisonings, car wrecks, falls and several chronic diseases, including coronary artery disease, hypertension, chronic obstructive pulmonary disease, chronic pain, behavioral health disorders, addictions and poverty . OverallThis important study was followed by another pivotal study, which demonstrated a dramatic 30% increase in cellular aging at a median chronological age of 30 years using a late-generation epigenomic clock based on DNA methylation, demonstrating the acceleration of the aging process from cannabinoid exposure in somatic (non-germ cell) tissues utilizing ‘state-of-the-art’ technologies .However, arguably of greater concern, is that fact that the nuclear blebs and bridges and chromosomal breaks and translocations well-described for sperm and oocytes in the experimental cannabis literature ,167 are Central to assigning a potentially causal relationship between the exposure and observed epidemiological effects was the presence of the plausible biological mechanisms of action . The pasWhile these many different modalities were explored in a number of previous publications and reviews ,101,102,Many layers of epigenomic regulation are described and the list appears to be rapidly increasing. While they may be listed individually, they are not independent and are coordinated across the various layers. The key parameters include DNA methylation, the histone post-translational modifications, various short and long non-protein coding RNAs, over 100 post-transcriptional modifications to the RNAs, including m6-adenosine RNA methylation , the position with respect to the nuclear lamina (which is suppressive of the gene transcription), the chromatin state (euchromatin or suppressive heterochromatin), the presence within the transcriptional factories of the topologically defined domains (and their controlling boundary elements) and the presence of the tethering elements  .It is now well understood that the cell lineage specification  is controlled epigenomically. This issue was first formalized in the epigenetic valley hypothesis of Conrad Waddington . It is aFor a long time, the mechanisms of aging were not understood and many competing and often complimentary and overlapping theories were advanced . HoweverThis implies that the epigenomic state is central and pivotal to the control of cancer, cell development and aging, which are the three principal themes of the present discussion.A recent detailed epigenome-wide association study (EWAS) by Schrott and colleges investigating the DNA methylation changes of human and mouse sperm both in cannabis dependence and withdrawal provides a 359-page Supplementary Appendix listing the detailed methylation changes . These rThere was widespread disruption between the main readers, writers and erasers of the epigenetic code. Hence, there were five hits for the DNA methyltransferases which added the methylation mark to the CpG islands and one hit for TET1 (ten-eleven translocase) which began the process of removing it. There was one hit for telomerase which controlled the end length of the chromosomes, and thus protected them against aging, three hits for polycomb repressors, five hits for the chromatin remodelers  and three hits for the UHRF (Ubiquitin-like with PHD and ring finger domains) family which controlled both DNA methylation and histone methylation and integrated the signaling in the two classes of pathways.There were 161 hits for the histone methyltransferases with methylate histones and 199 hits for the histone demethylases that remove this mark. There were eleven hits for both the histone acetyltransferases, which acetylate histone tailed and thereby made the genome accessible to the transcription machinery, and eleven hits for the deacetylases which removed this mark.Considering the key stem cell induction factors identified by Yamanaka, Oct3/4, Sox2, Klf4 and Myc , all fouAs noted above, chromosomal disorders hold a prominent place in the patterns of cannabinoid-related carcinogenic and teratogenic disorders. Indeed, when the length of all the chromosomes involved was summed (omitting duplications), it was found that an impressive 1765 megabases of the 3000 megabases of the human genome were directly implicated in the cannabinoid-related genotoxic disorders, which is 58.8% of the human genome. For this reason, the epigenomic findings of the Schrott EWAS dataset were of immense importance.Additionally, the rays of the mitotic spindle were composed of microtubules of polymerized tubulin. There were 106 hits in the Schrott EWAS database for tubulins. Importantly, tubulin undergoes numerous post-translational modifications which are thought to govern its intracellular trafficking and organellar addressing ,190,191.The centromeres are the critical central portions of chromosomes where binding occurs to the mitotic spindle. Centrosomal protein A (CENPA) was a modified version of histone 3 (H3) and CENPA replaced H3 in the centromere, which marked the location of the centromere. Upon this CENPA basis, a complicated scaffold of 17 proteins assembled which then was bound to the microtubules of the spindle via other kinetochore scaffolding proteins ,195,196.The proteins which cohere the ends of the human oocyte meiotic spindle so that two (and only two) spindle poles are formed, guiding the formation of two (and only two) daughter cells, are called centrosomal organizers. There were three hits for these proteins, including the nuclear mitotic apparatus protein (NUMA).The motor proteins which actually move the chromosomes along the microtubules towards minus end of the microtubules and the spindle poles after the anaphase checkpoint is released are called dynein motors which are controlled by a binding partner known as dynactin. There were seven EWAS hits for dynein–dynactin. Interestingly the intracellular kinesin motor moved protein and other cargo in the opposite direction towards the positive end of the microtubule and 218 hits were recorded for kinesin motors.Sumoylation was shown to be a key post-translational modification of the key proteins, which organizes the chromosomes and are known as the remodelers of the structure of the chromatin (RSC) complexes in yeast . SumoylaIt was also noted that the EWAS screen showed nine hits against RAD51, which was the key member of the high-fidelity homologous recombination (HR) pathway, but only one hit against RAD52, which was part of the low-fidelity non-homologous end-joining pathway . It was Thus, these many results clearly impacted and disrupted all the major functions of the chromosomes and likely provided a powerful epigenomic underpinning for the epidemiologically observed carcinogenic and teratogenic pathophysiology. Moreover, the DNA breakage was shown to be a prominent feature of the cannabis exposure of oocytes, sperm, lymphocytes and many other cells, and these finding imply that these lesions were preferentially repaired low-fidelity rather than high-fidelity pathways due to the epigenomic dysregulatory mechanisms.The Schrott EWAS study  revealedThere were ten hits each on neurexin and neuroligin, which are a ligand–receptor pair that mediate the receptor formation and scaffolding. There were eight hits on discs large homolog-associated protein 2 (DLGAP2), which is a protein known to be involved in synaptic scaffolding and the previously implicated ion autism development . SimilarRecent studies have shown that the massive overgrowth of the human cerebral cortex relative to other species is controlled by signaling between the Slit-Robo ligand–receptor pair ,200,201.Another key study found that the very high gradient of retinoic acid at the frontal pole of the forebrain was responsible for driving the frontal lobe outgrowth . The graThese data showed that the cannabinoid stimulated epigenomic pathways disrupted the synaptic processing across a broad range of receptor subtypes, synaptic scaffolding using several routes, and neural progenitor and forebrain outgrowth by inhibiting several of the main pathways responsible for this key proliferation action. Such findings indicated that mental illness and congenital neurological conditions, including autistic spectrum disorders and developmental disorders such as microcephaly and anencephaly, were more likely, as observed in an increasing number of large epidemiological studies on community cannabis exposure ,29,40,45This was, in turn, systemically important as brain aging has been well demonstrated to drive whole organism aging ,166,206.The issue of vascular aging has broader implications than simply the cardiovascular system since it has been aphoristically said that “you are as old as your arteries” ,212,213.Cannabis exposure has been shown to advance human cardiovascular age in an ecological longitudinal study .The key genes in arterial development are sonic hedgehog (shh), the vascular endothelial growth factor (VEGF) and notch and ephrinB2 signaling .Importantly, when investigating the EWAS-identified epigenetic methylation changes to human sperm, sonic hedgehog signaling was shown to be disrupted by nine hits on both the patched co-receptor and elsewhere, in addition to 185 hits on the key Gli3 transcription factor which signaled to the nuclear genome . Notch, The point of these findings was not only to identify that cardiovascular development can be disrupted in these ways but that the induced arterial aging can also induce the system-wide whole organism aging processes through an impairment of the stem cell quiescence/multiplication balance both directly and indirectly.The Schrott EWAS contained 73 hits for central nervous system dysfunctions, including the brain, neurological, synaptic, cerebral, neuronal and eye derangements.At total of 29 hits were noted for cardiovascular disorders, including the heart, atria, ventricles, atrioventricular valves and vessels.Additionally, 22 hits were noted for orofacial genetic lesions, including the head, sensory organs, palate, nose, anterior eye and ear derangements.Six hits were identified for limb development directly. Further exploration of a more exhaustive list of the limb morphogens revealed 130 hits for most of the key morphogens involved in limb and digit development, including the fibroblast growth factors (FGFs), retinoid signaling, Wnt signaling, bone morphogenetic pathway signaling and five genes from the sonic hedgehog (shh) pathway, namely MEGF8 (multiple EGF-like domains 8), TMEM107 (transmembrane protein 107), Gli3 (GLI gamily zinc finger 3), CHD7 (chromodomain helicase DNA-binding protein 7) and the patched receptor cofactor. Indeed, 185 hits for the key shh transcription factor Gli3 were found in the Schrott EWAS.There were 37 hits for development of the gastrointestinal tract, including references to the esophagus, large intestine, liver and pancreas. This epigenomic pattern was noted to be consistent with the pattern of the anomalies observed in the population cannabis exposure data from both the US and Europe ,36,40,51There were 23 hits observed for the urinary system, including the kidneys. When a more detailed exploration of gene regulation guided by a recent developmental renal cell map was used as a guide for data mining , a totalA total of 15 hits were identified for the body wall and embryo.Additionally, 60 hits were noted for the general otherwise unclassified disorders, including embryonic growth, DNA, mitochondria, microtubules, body trunk, body axis, ovarian reserve, breast disorders, granulocytes, myogenesis, vertebral growth and bone development.Hence, an abundance of epigenomic evidence exists to explain the broad spectrum and high severity of the teratological patterns observed in the many jurisdictions described. These findings are described in further quantitative detail elsewhere .The Schrott supplementary file lists 487 hits for “cancer”, 112 hits for “tumor”, 126 hits for “carcinoma”, 36 hits for “neoplasm”, 32 hits for “leukemia” and 17 hits for “lymphoma”. This totals 810 hits for cancer and its synonyms, making this one of the standout and major findings of this EWAS report.The report specifically mentioned many leukemias, lymphomas, myeloma and tumors of the breast, ovary colorectum, thyroid, liver, brain, pancreas, melanoma, stomach, esophagus and upper aerodigestive tract.As noted, all of these cancers have been described in association with cannabis exposure in historical ,227,228 By listing over 30 cancers by name and the genes whose epigenomic modulation was linked with them, these results provided a powerful pan-cancer mechanistic contributory explanation for the patterns of cancer epidemiologically observed in human populations.It is also worth noting that many of the more recent epidemiological reports proceeded beyond the methodologies commonly adopted in observation cohort studies ,229,230.A concise overview and introduction to aging was provided in the preceding sections. Since DNA methylation was shown to be a key determinant of the progressive decline of the function which characterizes the aging process, and since cannabis dependence and withdrawal was shown to widely disrupt both DNA methylation and demethylation and the histone code with which it is coordinated, the disruption of the aging process itself is not unexpected. As noted above, this was confirmed in somatic tissues experimentally and found to be of high magnitude at 30% at 30 years of age .p = 2.82 × 10−6 and a multiplicity-corrected p-value of 0.01258 [p = 1.18 × 10−5, multiplicity-corrected p-value = 0.02278) and this is the main counterbalancing force to the promoter hypermethylation of aging, it is easy to understand how the accelerated aging process is not only established initially, but how it might become a positive feed-forward process with time as age-related epigenomic changes are predisposed to further pro-aging epigenomic processes.It was also shown that cannabinoids can reduce the telomerase activity in a rat hepatocarcinogenesis model . The Sch 0.01258 . Indeed,Two of the key tissues with which we were concerned in the present context were the male and female gametes. It was understood that none of the presently available epigenetic clocks were suitable for the application to measure the relatively very hypomethylated ages of the gametes. However it does stand to reason that it may be possible to develop such an algorithmic clock mathematically. What the negative ages might mean, as they may relate to ages prior to birth, has yet to be determined biologically. Hence, it was not possible to measure gametal age directly or epigenomically at the time of writing.However, it was emphasized that the well-described presence of the characteristic aging nuclear changes on the sperm of the DNA chromosomal breaks and translocations  and oocyThere are various strengths and limitations to the present conceptualization. The strengths include the remarkable consistency across the many epidemiological studies, which clearly demonstrates the genotoxic harms of cannabis exposure in several different international jurisdictions, in relation to both the congenital anomalies ,80,81,82The major limitation of the present work is its preliminary nature in that we present an introductory conceptual framework which needs to be filled out and completed by numerous further laboratory studies. The purpose of the present paper is merely to draw attention to this remarkable concordance of cross-disciplinary results and indicate to researchers in the basic sciences that the field is ripe for detailed exploration in many studies with far-ranging consequences. In the same way that Harvard researchers were recently able to advance cellular and organismal age forwards and backwards by the induction of just a few DNA breaks and then demonstrate their phenotypical rescue with the OSK Yamanaka stem cell factors  so too mMoreover, the 810 differentially methylated genes implicated with cancer by the work of the Murphy lab  have notThe important questions that need to be considered include the following.How is the landscape of the post-translational histone modifications changed by cannabis exposure, particularly in relation to the key modifications, such as histone acetylation, which reliably mark active genes?How are the critically important issues of the modulation of super-enhancers and super-anchors affected by cannabinoid exposure?Can tissue de-differentiation be demonstrated experimentally from cannabinoids and, if so, which tissues are the most susceptible? What are their time course? What are their dose-response effect? In particular, in the ovaries, testes, brain, liver, heart, respiratory tract and immunocytes. Is this de-differentiation premalignant? Do cannabinoids induce premalignant field changes and cancerization from adverse genomic (DNA breaks), epigenomic or metabolomic effects or some interaction between all of these and more?Oocytes are particularly genomically fragile and non-renewable cells and have a very long life of many decades. Their genomic, metabolic and epigenomic vulnerability to cannabinoids needs to be characterized in detail. What are the effects of cannabinoids on oogenic stem cells both prenatally and postnatally?Since cannabinoids affect the mitochondria and cell metabolism adversely and this is closely related to maintenance of both the genome and epigenome, how does this compare to the Warburg effect observed in stem cells and cancer cells ? How is Such studies would provide an invaluable window of insight into the way the metabolome and epigenome are coordinated and bidirectionally co-regulate each other.In particular, the effect of lactylation as a key post-translational modification of the key metabolic enzymes and histones needs to be quantified, as this has also been shown to be a critical post-translational modification for stem cells, cancer cells and cancer stem cells with potentially far-reaching and cross-disciplinary applications in aging medicine and cancer biology ,239.As described, cannabis widely disrupts many of the key enzymes of the epigenetic machinery itself. What are the implications of this?As discussed, cannabinoids disrupt key elements of the mitotic spindle, microtubule physiology, spindle pole formation and kinetochore and centrosome formation. These are very far-reaching findings as chromosomal mis-segregation has implications in many fields, including male and female fertility optimization, sperm and egg storage, the preservation and fertility of medicine, the biology of aging, cancerogenesis, congenital anomalies in early pediatrics and neurodevelopmental alterations in later pediatrics. This field has been largely ignored but is very much in need of detailed exploration and explication at the cellular and molecular levels from the point of view of the impacts of cannabinoids.As described by epigenomically disrupting both CTCF and the major components of the cohesion complex, it seems inevitable that cannabis must disrupt the basic machinery of chromatin looping and gene expression itself. What are the implications of this potentially very far-reaching derangement?What is it about testicular germ cells that makes them particularly susceptible to cannabinoid oncogenesis? Is it the DNA hypomethylation of germ cells to start?Techniques are emerging to allow the characterization of the non-protein coding, regulatory and repeat sequences in the genome. In what way is this normal physiology perturbed by cannabinoid exposure?DNA hypomethylation of the gene bodies and gene deserts is likely to result in the mobilization of transposable elements in the genome, which precipitates genomic instability, cytoplasmic and extracellular immune activation  and, thus, aging and oncogenesis in both the immune and genomic pathways. This field needs to be explored and further developed.The advancement of human epigenomic age from cannabis exposure at a single age has been mentioned . What isMany different cannabinoids need to be profiled in vitro to characterize their multichannel epigenomic effects  in the modern era to define the breadth of their epigenotoxicity as a possible or likely class effect.The well-documented exponentiation of the cannabinoid dose-response effects in many cellular and metabolic assays needs to be formally explored in the modern epigenomic context and its public health significance needs to be carefully considered.These and a host of similar questions are now present for careful review and exploration by modern laboratory techniques with almost certainly far-reaching implications for both the basic sciences and clinical medicine.The above overview of several large datasets is notable for many reasons. There is a surprising uniformity in the findings from the three domains of cannabinoid-related teratogenesis, carcinogenesis and the acceleration of aging processes, which together imply a clinically significant if widely underrated cannabinoid genotoxicity. Moreover, the severity of the findings in all three domains is considerably greater than is commonly supposed.The other very striking feature is that the predictive power of the epigenomic profiles provided by the recent epigenome wide studies, particularly that of Schrott and colleagues , is the The wide spectrum of cancers is specifically explained and the organ-specific nature of many congenital anomalies is explained. The pathophysiology of the wide spectrum of chromosomal disorders which occur as both malignant disorders and congenital anomalies is also elegantly explained by several mechanisms. The pattern of limb anomalies, including syndactyly and polydactyly, is explained by the widespread epigenomic disruption of most of the key limb morphogen gradients. Most particularly, the patterns of accelerated aging are explained by the epigenomic inhibition of DNA methylation, demethylation and telomerase, among others, and confirmed by the recent pattern of chronic illness studies and epigenomic clock investigations.Most concerning, the very real prospects that both sperm and oocytes are epigenomically aged prior to conception carry severe and dire prospects for public health and multigenerational epigenomic transmission and beg a formal assessment using gamete-appropriate epigenomic clocks whose development must be considered a major research priority.With the increasing interest in the pharmaceutical use of cannabis and cannabis derivatives for medical management, it is increasingly important that the balance between the therapeutic risks and benefits for each potential cannabis/cannabinoid application is thoroughly understood. The mandate of regulatory global authorities is to not succumb to strong commercial and popular interest in the promotion of cannabis/cannabinoids until the risk–benefit appraisal can be accurately undertaken for each potential cannabis application, considering, among other items, the potency, severity of disease, stage of human development and duration of use.


Purposes: A stent-assisted coil (SAC) is a safe and effective treatment modality for some complex intracranial aneurysms, especially for wide neck aneurysms. However, some wide neck aneurysms with a tough angle and located in the posterior communicating artery (PCoA) are challenging to treat with a SAC. This study aimed to examine and discuss the swinging-tail technique for treating wide neck aneurysms located in the PCoA using a SAC by Prof. Lv. Materials and Methods: We retrospectively reviewed our institutional clinical database and identified nine patients with neck aneurysms located in the PCoA, and these patients underwent the swinging-tail technique by Prof. Lv, which is a novel technique of releasing a stent, from June 2016 to September 2021. Results: In this study, nine patients underwent SAC treatment using the swinging-tail technique by Prof. Lv. Aneurysmal complete occlusion was observed in every patient without any complications, as shown by immediate postoperative angiography. Additionally, the modified Rankin scale was monitored for clinical outcomes in the follow-up. One patient died postoperatively due to severe SAH with an intraventricular hemorrhage. Four of nine patients underwent imaging follow-up, demonstrating the complete occlusion of aneurysms; eight patients underwent clinical follow-up and achieved a favorable clinical outcome . Conclusion: The SAC treatment for wide neck aneurysms located in the PCoA can be challenging for operators because of the specific location, resulting in inadequate vessel wall apposition by antegrade stenting via the ipsilateral vessel. In this circumstance, the swinging-tail technique may be a feasible and effective choice. Endovascular stents are widely used during the embolization of intracranial aneurysms. Theoretically, stent deployment provides a mechanical scaffold for coils inside an aneurysm, allowing improved packing density and neck coverage . When trThis retrospective study was reviewed by Beijing Tiantan hospital’s institutional review board and approved by the ethics committee of Beijing Tiantan Hospital. The patients provided their written informed consent to participate in this study.In this study, we retrospectively reviewed the patients diagnosed of aneurysms located in the PCoA—confirmed by digital subtraction angiography (DSA) and/or computed tomography angiography (CTA)—and underwent endovascular treatment by Prof. Lv, who has sufficient clinical experience in the treatment of intracranial aneurysms using SACs, from June 2016 to September 2021. Initially, according to preoperational multidisciplinary discussions, preferences for microinvasive procedures were conveyed by the patients’ relatives; for the native characteristic of a wide neck aneurysm, endovascular treatment by SAC was regarded as the first line of treatment for the patients. Furthermore, nine patients who underwent SAC treatment using the swinging-tail technique by Prof. Lv were recruited for this study.Patients without a hemorrhage were premedicated with a dual antiplatelet regimen  for 5 days before treatment. Patients with a hemorrhage were administered a loading dose of 300 mg of aspirin and 300 mg of clopidogrel, 2 h preoperatively. Intraoperatively, an intravenous bolus dose of heparin (100 IU/kg) was administered. Heparinization was continued to maintain an activated clotting time two- to three-fold greater than the baseline value throughout the procedure. Dual antiplatelet therapy was continued for 3 months postoperatively, and aspirin was continued for life, to prevent the formation of thrombi in the stents.By performing the precise preoperative evaluation of aneurysmal characteristics according to DSA and/or CTA in detail, we found, differentiated from the aneurysms located in internal carotid artery (ICA) C7 segment, that the aneurysms admitted to our study primarily affect the PCoA rather than the ICA C7 segment. Furthermore, there is an acute angle between the PCoA affected by an aneurysm and the proximal segment of the ICA, which limits the choice of conventional endovascular treatment by a SAC , because of inadequate wall apposition. In this circumstance, SAC via the contralateral ICA and anterior communicating artery (ACA) may be an available choice . HoweverAll procedures were performed with the patient under general anesthesia via the transfemoral approach, and systemic heparinization was administered after the placement of a sheath. A suitable guiding catheter was navigated to the target location via a feasible procedural course. Three-dimensional rotational DSA was performed to determine the best working projection and to measure the size of the aneurysms, the diameter of the ICA and PCoA, the angle between the PCoA affected by aneurysms, and the proximal segment of the ICA (below the ostium of the PCoA). Suitable stents, with a high metal coverage rate in diseased vessels, were selected according to the aneurysmal measurements, the diameter of the PCoA, and the proximal segment of the ICA, with the aim of achieving complete coverage of the aneurysmal neck and a good wall apposition between the stent, PCoA, and the segment of the ICA. We used suitable coils to completely occlude the aneurysmal sac with the assistance of stents.The points of the swinging-tail technique are as follows: (1) A suitable stent was selected according to the precise aneurysmal measurements, the diameter of the PCoA, and the proximal segment of the ICA, as assessed by intraprocedural DSA. A suitable stent is of importance for reducing the possibility of aneurysmal recurrence A; (2) A The key point is that a suitable length of the stent segment released in the ipsilateral ICA was estimated; this was important because an unstable stent situation in the blood flow was ascribed to a short length of stent segment, and the failure of the swinging-tail technique was ascribed to a long length of stent segment.This case series included nine patients with nine aneurysms who underwent the swinging-tail technique , with a mean (±standard deviation) age of 62.8 ± 9.7 years . Two patients were diagnosed with a subarachnoid hemorrhage (SAH) by preoperative computed tomography (CT). Nine aneurysms were located in the PCoA, as confirmed by preoperative DSA. The patients’ demographics and aneurysmal characteristics are shown in In this study, all patients tolerated the procedure well, without any intraprocedural complications. Nine patients with nine aneurysms underwent endovascular treatment (SACs) using the swinging-tail technique by Prof. Lv. Additionally, all patients successfully underwent SACs using the swinging-tail technique . To reduClinical follow-up data were available for all of the patients  by enrolling the patients or communicating via telephone. In our study, we selected the modified Rankin scale (mRS) score as our standard criteria for evaluating the clinical conditions of every patient. Preoperatively, two  patients were diagnosed with SAH by preoperative CT; they had relatively poor clinical conditions, with mRS scores of two and five, respectively. After undergoing treatment, three  patients showed improvements in their clinical conditions, three patients  were diagnosed with an intracranial aneurysm by physical examination coincidently without any associated symptoms. Two  patients experienced no clinical improvement and were presented with a chronic headache associated with the severe stenosis of dual vertebral arteries and cervical spondylosis, with an mRS score of one during follow-up. One patient  died of severe SAH with a mass intraventricular hemorrhage three days after the operation. The detailed clinical follow-up outcomes are shown in Four patients underwent a follow-up with DSA . Generally, we selected the final DSA follow-up of every patient as the time point used to evaluate the efficacy of the endovascular treatment. Four patients achieved the complete occlusion of aneurysms, as confirmed by the results of the DSA follow-up. DSA follow-up data are shown in Currently, endovascular embolization is recommended as the first choice for the treatment of most intracranial aneurysms due to the advanced technique associated with endovascular embolization; this is especially evident for the application of the stent-assisted coiling technique, combined with the advantages of minimal invasiveness and a quick recovery . Stents Because these lesions are rare and differentiated from the aneurysms of the posterior communicating segment of the distal ICA, their anatomical relationships have been poorly described. Similarly, reports concerning the endovascular treatment of these lesions are rare, and the technical aspects are poorly detailed . AneurysIn our study, we found that all aneurysms were wide-necked aneurysms (neck diameters >4 mm or a dome-to-neck ratio <2), and the angle between the PCoA affected by aneurysms and the proximal segment of the ICA was acute. When evaluating the technical feasibility, Prof. Lv, an experienced neuro-interventional expert, believed that conventional endovascular treatment  would not be able to achieve excellent therapeutic outcomes, with a large possibility of recurrence. Recently, for this complex lesion, Yang et al. reported the novel proximal-end stenting technique, which uses the proximal end of the stent to assist with the embolization of a wide-necked aneurysm located in the PCoA . HoweverAccording to the preoperative DSA performed in every patient, we initially rejected the conventional endovascular treatment modality  because of the particular location of the aneurysm and the acute angle between the PCoA and the proximal segment of the ICA. Yang et al.  reportedAdditionally, although these lesions could not be effectively managed by conventional antegrade stenting via the ipsilateral vessel, we developed a novel technique via the ipsilateral vessel (the swinging-tail technique) to treat these complex lesions. The swinging-tail technique is a procedure where the stent is released from the PCoA affected by an aneurysm to the distal segment of the ICA via the ipsilateral ICA. In this situation, the distal end of the stent is located in the PCoA affected by an aneurysm, and the proximal end of the stent is located in the distal segment of the ipsilateral ICA, at the ostium of the PCoA. The primary advantage of this technique is that the aneurysmal neck is completely covered by the stent, and the ipsilateral PCoA is simultaneously preserved to the utmost extent, with a low possibility of recurrence and ischemic events. Fortunately, in this study, all patients obtained excellent clinical outcomes according to the results of the clinical follow-up, except the patient with a poor preoperative condition who died three days after treatment due to a severe SAH with a mass intraventricular hemorrhage; all aneurysms that underwent the treatment of the swinging-tail technique achieved complete occlusion intraoperatively and four aneurysms had no signs of recurrence according to intraoperative DSA and DSA follow-up, respectively. Similarly, retrograde stenting via the contralateral vessel can accomplish the same therapeutic outcome as the swinging-tail technique in the condition of satisfying the anatomic vascular structure. The swinging-tail technique requires a greater technical ability for operators so that it is not regarded as a normal treatment modality for this kind of refractory lesion. However, according to our treatment strategies for the aneurysms located in the PCoA, the swinging-tail technique can be an alternative treatment modality.This study has several limitations. First, this was a retrospective study, with a relatively small number of patients. Second, the swinging-tail technique, as a novel technique for treating aneurysms located in the PCoA, requires the increased technical ability of operators. Therefore, popularizing this technique for the treatment of the aneurysms located in the PCoA will be difficult and time-consuming, which precludes the precise therapeutic evaluation of this technique for these complex lesions. Third, although our patients underwent endovascular treatments—with complete occlusion of the aneurysms, as confirmed by immediate postoperative angiographies—long-term follow-ups remain necessary, and we only received the DSA follow-up of four patients. Therefore, a long-term study involving a large cohort is required to obtain sufficient evidence to evaluate the effectiveness and safety of the swinging-tail technique for the treatment of aneurysms located in the PCoA.An aneurysm located in the PCoA is challenging for SAC treatments because of the specific location between the PCoA and ICA. The PCoA affected by aneurysms and the proximal segment of the ICA form an acute angle, which makes the apposition of stents for an aneurysmal neck insufficient, resulting in increasing the possibility of the recurrence of aneurysms. In this circumstance, the swinging-tail technique, via the ipsilateral vessel, is a feasible and effective choice, though requiring the increased technical ability of operators. To evaluate the effectiveness and safety of the swinging-tail technique, a long-term clinical and/or DSA follow-up is required for these patients.


Advancements in molecular technologies have extended the potential role of circulating biomarkers in diagnosis, disease monitoring, and outcome prediction. Research is working towards personalized treatment in early breast cancer patients, and tools for intensification/de-intensification strategies are needed to guide clinicians’ choices. In this review, we explore the current developments of promising biomarkers in early breast cancer for cancer detection and tailoring neoadjuvant therapies.The research on non-invasive circulating biomarkers to guide clinical decision is in wide expansion, including the earliest disease settings. Several new intensification/de-intensification strategies are approaching clinical practice, personalizing the treatment for each patient. Moreover, liquid biopsy is revealing its potential with multiple techniques and studies available on circulating biomarkers in the preoperative phase. Inflammatory circulating cells, circulating tumor cells (CTCs), cell-free DNA (cfDNA), circulating tumor DNA (ctDNA), and other biological biomarkers are improving the armamentarium for treatment selection. Defining the escalation and de-escalation of treatments is a mainstay of personalized medicine in early breast cancer. In this review, we delineate the studies investigating the possible application of these non-invasive tools to give a more enlightened approach to escalating/de-escalating strategies in early breast cancer. Early breast cancer (eBC) is considered a potentially curable disease; however, nearly 30% of cases will evolve into metastatic disease even after a long follow-up . DefininLiquid biopsy is the most intriguing resource for circulating biomarkers such as cell-free DNA, circulating tumor DNA (ctDNA), circulating tumor cells (CTCs), and circulating tumor RNA (ctRNA). Furthermore, the peripheral inflammatory cells may easily be evaluated as non-invasive biomarkers from a simple blood sample, revealing information even at the early stages of the diagnostic–therapeutic approach . Other iIn the preoperative setting of eBC, individualizing predictive response factors to neoadjuvant treatment and the achievement of a pathological complete response (pCR) is still an unmet need. PCR is increasingly being considered as a potential surrogate for survival outcomes, but there is still a lack of evidence to support its connection with different survival endpoints . As a coSimilarly, in triple-negative (TN) disease, the use of neoadjuvant pembrolizumab in association with standard chemotherapy has been shown to increase the possibility of pCR achievement . HoweverIn the view of personalized care, it is clear there is a need for useful tools to predict treatment efficacy and prognosis before surgery and presurgical treatments to improve the therapeutic strategy. New algorithms have been developed to extrapolate the prognostic and predictive information provided by tissue biomarkers such as genomic tests prospectively validated in the adjuvant setting , and others are under development  . In partCirculating inflammatory cells are involved in the process of metastasizing and seem to also have a role in the first phase of cancer promotion and progression, as observed from studies on metastatic BC patients ,19,20. IPretreatment values of the neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and monocyte-to-lymphocyte ratio (MLR) may reveal predictions for treatment efficacy, pCR, and survival outcomes. A meta-analysis including more than 17,000 patients with early and advanced BC revealed an association between high pretreatment NLR and a poorer prognosis in eBC . The ideElevated platelet-related markers may also be associated with a poor prognosis in eBC. The platelet counts and platelet volume indexes seem to represent prognostic factors in eBC as indicators of an increased risk of metastasis . In addiMonocytes are particularly involved in the process of metastasizing, favoring cancer cell dissemination even in the very early stages, and managing the development of the metastatic niche . The depThe development of lymphopenia after NACT was not associated with pCR achievement in a retrospective analysis regarding exclusively TN eBC . HoweverAdditionally, the preoperative systemic immune-inflammation index  showed to potentially give prognostic information before surgery . A high Certainly, the retrospective nature of the majority of the data available and the retrospective findings of ongoing studies (NCT05468710) will be a clear limitation for the clinical utility of inflammatory indexes in terms of prognostic validity. Prospective studies are awaited to thicken the body of evidence about the prognostic value of immune circulating cells in eBC.CTCs are cancer cells that are detectable in the bloodstream . CTCs ha® . W. W187].  ongoing .The search process according to the PRISMA Guidelines. * 20 studies considered for The intriguing results of novel circulating biomarkers will be useful in evaluating more precisely the prognosis of eBC; however, their use has not been established yet in clinical daily practice . FurtherCurrently, not all the assays are equal to the others. Too many differences between the detection techniques do not allow for a defined, univocal method and consequently, we are far from a definitive conclusion regarding which test has the most predictive value for the clinical outcome. Accordingly, the clinical utility for cancer screening, response monitoring, and evaluation of treatment efficacy of circulating biomarkers is not completely defined and not applicable except in clinical trials. The gap between bench and bedside can be filled when circulating biomarkers are used regularly in wide prospective trials on neoadjuvant approaches in eBC, establishing their direct role in the decisional process. Results of ongoing trials are awaited to improve tools for escalating and de-escalating approaches in eBC. The combination of clinical, pathological, genomic, laboratory, and radiologic findings will contribute to a more precise stratification of the patients and the diseases. The development of predictive models and nomograms with the combination of multiple features should be the future challenge to reduce the need for invasive approaches.


China has undergone rapid economic growth and diet transition over the past decades, along with environmental challenges. To help to achieve carbon peaking by 2030, this study investigates the time-variable diet-related greenhouse gas emissions (GHGEs) from 2000 to 2020 and examines the responses of food demands and diet-related GHGEs to an increase in per capita income and animal-based food prices. Based on the Almost Ideal Demand System model, addressing the problem of the endogeneity of food expenditure, we estimate the demand elasticities for six food groups and analyze their changing trends with time. Then, we perform two projections. One is to project the impacts of income growth on food demands and diet-related GHGEs, taking into consideration dynamic income elasticities as per capita income increases; the other is to project the effects of an increase in prices of different animal-based foods on food demands and diet-related GHGEs. Our results reveal that diet-related agricultural GHGEs show a continued increase in the short term, but the increased effect tends to decline gradually. The dominant driver of increased diet-related GHGEs is the increased consumption of beef, mutton, and pork. However, an increase in prices of beef and mutton; pork; and poultry and eggs can reduce diet-related GHGEs, while a decrease in the prices of aquatic products can also deliver a significant decrease in GHGEs. The results imply that policymakers should take an interest in the environmental impacts of diet transition and promote a more sustainable diet structure. For example, crop and livestock production within the farm gate contribute more than 50% of the total anthropogenic methane (CH4) and 75% of the nitrous oxide (N2O) emissions [The agricultural sector is an important contributor to anthropogenic greenhouse gas emissions (GHGEs) . GHGEs fmissions ,4. BesidIn addition to the mitigation strategies in food supply chains, dietary patterns are also critical to mitigating agricultural GHGEs through their impact on supply activities. Many studies have explored a series of approaches to reduce agricultural GHGEs from the supply side, such as technological innovation and production management . From anChina is an interesting and important case in regards to this problem. First, China is a traditional agricultural country, which feeds about 20% of the global population. The agricultural GHGEs account for 17%  of the total anthropologic GHGEs in China, and the ratio is higher than the average global level . This stA great deal of work has been done to evaluate diet-related GHGEs and propose strategies to reduce GHGEs. Previous studies have calculated the GHGEs from plant- and animal-based food  and evalMany studies estimated food demand elasticities using demand models or meta-analysis, some of which further projected the food demands in the future. These results showed that income responsiveness was higher for meat consumption, and meat demand would continue to increase with income growth ,24,25. HThis study aims to estimate the dynamic changes of food demand elasticities with time-series data, and project the responses of food demands and diet-related agricultural GHGEs to an increase in per capita income and animal-based food prices. This study fills the gap in the literature by (1) projecting the impacts of a rise in income and animal-based food prices on diet-related GHGEs considering the change in demand elasticities; (2) analyzing the change trends in demand elasticities with time-series data. We employed the data from the China Statistical Yearbook (2001~2021) and divided the residents’ consumed foods into six groups. First, we clarified the food demand characteristics and related agricultural GHGEs from 2000 to 2020. Then, we used the Almost Ideal Demand System (AIDS) model to estimate the demand elasticities of six food groups and analyzed their dynamic changes. Last, we performed two projections. One is to simulate the impacts of income growth on food consumption and diet-related GHGEs considering the dynamic changes of income elasticities; the other is to project the responses of food consumption and diet-related GHGEs to an increase in prices of different animal-based foods.The remainder of the paper is organized as follows. The conceptual framework of this study is shown in For different agricultural products, the differences in greenhouse gases released in agricultural production processes are substantial. Generally, the carbon emission intensity per unit mass of animal-based food is much higher than that of plant-based food, especially for ruminant food such as beef and mutton . This meTo estimate the demand elasticities for various food groups, we employed the Almost Ideal Demand System (AIDS) model derived by Deaton and Muellbauer . The budIn addition to price and income, demographic structures also affect food demand . There and Robin  by usingstimator , and thestimator .The food expenditure and price elasticities can be derived by the following equations:Further, to measure the income elasticities of various food groups, we used Working’s model  to analyThen, the income elasticity of the total foods can be expressed as follows:The unconditional (income) elasticity for various foods can be derived by:This study mainly focuses on the direct and indirect GHGEs in the agricultural production process; the GHGEs released from storing, transporting, and cooking are not included. The carbon emission intensities of major food groups and their references can be found in According to the agricultural carbon emission intensity coefficients and per capita annual consumption of various food groups, the per capita agricultural GHGEs related to individual food and total food consumption over the years can be expressed as follows, respectively:ood item , and qkiThe data for per capita food consumption, disposable income, demographic structure, and the consumer price index come from the China Statistical Yearbook (2001–2021), and the food market prices from 2003 to 2020 come from the China Yearbook of Agricultural Price Survey (2004–2021). We used the proportion of residents aged 15 to 64 years as the demographic structure variable. Due to missing data, the food market prices before 2003 are derived by the consumer price sub-index and food market prices in 2003. Both per capita income and food expenditure have flattened the consumer price index based on the data from the year 2000. As the statistical criterion of urban residents’ grain consumption before 2012 and after 2013 is processed grain and raw grain, respectively, we followed the methods of Cao et al.  and usedSince the 21st century, China has experienced rapid growth in per capita disposable income and food expenditure . The perFurther, we analyzed the expenditure shares of each food group from 2000 to 2020 in China . Among pTo conserve space, only expenditure, income, and price elasticities every four years are shown in The own-price elasticities for vegetables, fruits, pork, beef, mutton, and aquatic products are negative and statistically significant, while those for staple foods, poultry and eggs are not sensitive to price changes . DiffereIn this section, we perform two projections. One is to project the response of food demands and corresponding agricultural GHGEs to a 1% increase in per capita income. This is based on income elasticities for individual food groups . The othBefore performing the first projection, we assume that the food prices are to be constant. According to the above results, the income elasticities show a downward trend as income increases; hence, here we perform three scenarios. S1 shows the average income elasticities from 2000 to 2020; S2 includes the average income elasticities from 2010 to 2020; S3 reduces the average income elasticities from 2010 to 2020 by 10%, according to the changing trend in income elasticities . Before 2eq under S1 and S2, respectively, while these decrease by 0.076 kg CO2eq under S3. This implies that the decreased income elasticities drive a lower increase in total diet-related agricultural GHGEs, and even reduce them.2eq. This is due to the substitute effects of staple foods, beef, and mutton. This result indicates that we might reduce GHGEs by raising the price of beef and mutton and cutting the price of aquatic products.Given the continued economic growth and diet transition, this study analyzed the time-variable demand elasticities and explored the responses of food demand and diet-related agricultural GHGEs to changes in per capita income and food prices. Consistent with the results of previous studies ,35, we fOur results show that income elasticities for most foods tend to decline, which are consistent with previous findings, although the exact elasticity values from various studies are different due to various data sources, food categories, and methods applied . The chaFurther, our projections show that the demand for all foods, except staple foods, will increase as per capita income increases. This is according to the finding of Zheng et al. , who revFrom the demand side, a single measure is also not enough to achieve carbon neutrality and mitigate the projected increase in environmental burden. In addition to improving diet, international trade and reducing food waste and food processing are also expected to deliver emission reductions ,6,43. FoThere are several limitations in this study, which should be further addressed in the future. First, the heterogeneities of regional disparity and the urban-rural gap should be further analyzed. Second, the data from the China Statistical Yearbook only include residents’ food consumption at home, ignoring the food consumption away from home. As meat demand, especially beef consumption, away from home grows more than proportionally to total food consumption , our resThis study investigated diet-related agricultural GHGEs in China and their responses to an increase in per capita income and animal-based food prices. The AIDS model, considering the issue of food expenditure endogeneity, was used to estimate demand elasticities for six food groups from 2000 to 2020. Then, the impacts of an increase in per capita income on food demands and diet-related agricultural GHGEs are projected considering the trend in the changes in income elasticities. Meanwhile, we also projected the responses of diet-related agricultural GHGEs to an increase in the prices of pork, beef and mutton, poultry and eggs, and aquatic products, respectively.Our results indicate that the food demand and diet-related agricultural GHGEs will increase in the short term, but the increased effect tends to decline gradually. In the long term, the total diet-related agricultural GHGEs might decrease due to a lower increase in animal-based food and a greater decrease in staple foods. The dominant driver of increased diet-related GHGEs is the increased consumption of beef and mutton and pork. However, increases in prices of beef and mutton, pork, and poultry and eggs can reduce diet-related agricultural GHGEs, while an increase in the price of aquatic products has the opposite effect.Along with remarkable economic development, China is also facing environmental challenges. On the road to achieving carbon peaking and carbon neutrality, this study has important implications for policymakers. With the continued increase in per capita income, although diet transition improves residents’ nutritional status, the government should pay attention to the environmental impacts driven by the rapid increase in animal-based food demand. In addition to the mitigation strategies from the supply side, the government can reduce GHGEs through food demand management from the demand side, while ensuring residents’ nutritional intake, for example, by subsidizing low-carbon food or taxing carbon-intensive food to promote a more sustainable diet structure.


Uric acid (UA) is the final product of purine metabolism in the human body, and impaired purine metabolism can increase the uric acid in serum, finally resulting in hyperuricemia (HUA). Current evidences suggest that urates might have antioxidant properties under certain circumstances, but most evidences suggest that urates promote inflammation. Hyperuricemia leads to the formation of urate crystals, which might be recognized as a red flag by the immune system. Such a response stimulates macrophage activation, leads to the activation of NOD-like receptor protein 3 (NLRP3) inflammasome vesicles, and ultimately the production and liberation of interleukin-1b (IL-1b) and interleukin-18 (IL-18), which can mediate inflammation, apoptosis and necroinflammation and cause an inflammatory cascade response. The kidney is one of the most commonly affected organs in HUA, which promotes the development of chronic kidney disease (CKD) by damaging endothelial cells, activating the renin-angiotensin system (RAS), and promoting inflammatory responses. Pharmacological interventions and lifestyle modifications are the primary means for controlling gout and lowering UA. The febuxostat is safe for CKD patients in the UA lowering therapy. Although dialysis can reduce UA levels, the application of drug is also necessary for dialysis patients. This article reviews the synthesis and metabolism of UA, etiology of HUA, the relationship between HUA and kidney disease, the treatment of gout and gouty nephropathy (GN). Uric acid is the final metabolic product of purines in humans. As the final product of exogenous purines from food and endogenous purines from damaged and dead cells, uric acid is synthesized mainly in the liver, intestine and vascular endothelium . The kidUnder physiological conditions, the synthesis and excretion of uric acid in our body is in balance. Hyperuricemia results when this balance is disturbed. Typically, levels of serum uric acid >6.8 mg/dl are considered to be hyperuricemia . The oveWhen the level of serum uric acid exceeds the solubility threshold, uric acid precipitates into crystalline urate crystals, which manifest as acute episodes of painful arthritis, forming gout . StudiesIt has been shown that urate has properties to scan for free radicals and has strong antioxidant capacity in human body . Uric acGouty nephropathy, also known as uric acid nephropathy, is a series of kidney disorders caused by an increase in uric acid in the human serum, which accumulates in the renal tubules and interstitium over a long period of time. Renal damage is a common comorbidity of gout and as kidney function declines, uric acid excretion through the urine is reduced, leading to hyperuricemia. Persistent hyperuricemia leads to the formation of urate crystals in joints and tissues . A recenThe mechanism of GN is mainly related to hyperuricemia and the deposition of monosodium urate crystals in the body. Survival conditions comprise high purine intake, excessive obesity and high dietary fructose concentration drinks, combined with the abuse of some drugs that affect the metabolic process of acid, such as thiazide diuretics, salicylates, and other metabolic substances such as lactic acid, ketone bodies and angiotensin. Excess uric acid deposited in the capillaries greatly increases the burden on the kidneys, leaving them in a state of long-term compensatory work, which eventually leads to a decrease in the filtration function of the kidneys and the deposition of urate crystals in the kidneys, causing lesions. Monosodium urate crystals precipitate in the renal tubules , causing acute gouty nephropathy. Uric acid stones may develop in 15-20% of patients with acute gouty nephropathy . ChronicThe main goal of gout treatment is to remove all urate crystals by lowering uric acid levels below 6 mg/dL. The process of deposition of urate crystals is reversible; crystals continue to form in gout patients and persist in patients with hyperuricemia, but dissolve when serum uric acid is lowered below the saturation point and the associated inflammation subsides with the disappearance of urate crystals . ImportaFor gout sufferers, dietary control is advocated. Reduce the consumption of high purine foods such as animal offal, red meat, sugar, seafood, soda and alcoholic beverages. Eat plenty of vegetables, vitamin C, skimmed milk, low-fat yoghurt, soy products, drink plenty of water (keep daily urine output above 2000ml), and avoid full meals. Alkalinize urine to pH 6.2-6.5 . Lifestyle modifications: exercise properly, lower body mass, drink green tea, maintain proper hunger (1 hour of hunger rests organs and increases longevity genes), promote early dinner, and in addition, educate patients properly to improve compliance and treatment outcomes.2 are reluctant to use allopurinol because of the potentially fatal allopurinol hypersensitivity syndrome (AHS) and the poor prognosis for patients with impaired renal function who develop AHS  but no reduction in renal function : XO inhibitors that inhibit uric acid synthesis, such as allopurinol and febuxostat, and drugs that improve the excretion of uric acid . Compared with allopurinol, previous studies have shown febuxostat to be more effective and safer. Recent studies have shown that ULT has no effect on the occurrence of the primary endpoint events . XO inhibitors have been reported to be beneficial in CKD. A meta-analysis showed that XO inhibitors reduced the risk of end stage renal disease significantly and also improved estimated glomerular filtration rate (eGFR) from data from randomized controlled trials with long-term follow-up (>3 months) . Furtherelop AHS . Febuxosfunction , which mfunction .Regarding the cardiovascular safety of febuxostat versus allopurinol in patients with gout and CVD, the use of febuxostat in CVD patients has been controversial. CARES conducted a large randomized controlled trial (RCT) in the United States in patients with gout and pre-existing cardiovascular disease. CARES randomly assigned 6190 patients with gout and cardio-vascular disease to receive febuxostat or allopurinol and patients were stratified according to kidney function. The CARES study found that all-cause and cardiovascular mortality were higher in the febuxostat group than in the allopurinol group  . FAST isDialysis provides appropriate clearance of serum uric acid. Relevant studies have shown that the average uric acid in hemodialysis patients is less than 5 mg/dL and the average serum uric acid after dialysis is less than 1 mg/dL, suggesting that the initiation of hemodialysis leads to clearance of tophus . It has Much of the current knowledge of the biological role of uric acid comes from experimental studies that have revealed that uric acid is associated with immune system activation and inflammation hyperuricemia may play a key role in the development and progression of CKD. Available evidence suggests that uric acid reduction therapy may slow the progression of CKD, although the molecular mechanism of uric acid-induced gout nephropathy remains to be further understood. Gout patients should be screened for renal function and clinicians should be aware of the link between gout and impaired renal function. The use of ULT drugs remain controversial. Long-term dietary control, lifestyle modification and patient education are the cornerstones of treatment.YM contributed to the conception and the writing of the article. BD performed the framework. ZG gave the constructive discussions to the article. LX revised important intellectual content critically for important intellectual content. 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.


Level of motivation, responsiveness to rewards and punishment, invigoration of exploratory behaviours, and motor performance are subject to daily fluctuations that emerge from circadian rhythms in neuronal activity of the midbrain’s dopaminergic system. While endogenous circadian rhythms are weak in the ventral tegmental area and substantia nigra pars compacta, daily changes in expression of core clock genes, ion channels, neurotransmitter receptors, dopamine-synthesising enzymes, and dopamine transporters, accompanied by changes in electrical activity, are readily observed in these nuclei. These processes cause dopamine levels released in structures innervated by midbrain dopaminergic neurons  to oscillate in a circadian fashion. Additionally, growing evidence show that the master circadian clock located in the suprachiasmatic nucleus of the hypothalamus (SCN) rhythmically influences the activity of the dopaminergic system through various intermediate targets. Thus, circadian changes in the activity of the dopaminergic system and concomitant dopamine release observed on a daily scale are likely to be generated both intrinsically and entrained by the master clock. Previous studies have shown that the information about the value and salience of stimuli perceived by the animal is encoded in the neuronal activity of brain structures innervating midbrain dopaminergic centres. Some of these structures themselves are relatively autonomous oscillators, while others exhibit a weak endogenous circadian rhythm synchronised by the SCN. Here, we place the dopaminergic system as a hub in the extensive network of extra-SCN circadian oscillators and discuss the possible consequences of its daily entrainment for animal physiology and behaviour. Historically, the suprachiasmatic nucleus of the hypothalamus (SCN) has been considered the only, master circadian clock which governs all rhythmic changes in physiology and behaviour occurring at a daily timescale. However, evidence accumulating over the last 2 decades questions the supremacy of the SCN . With thNeurons located in the ventral tegmental area (VTA) and substantia nigra pars compacta (SNc) have been suggested to possess such circadian clock properties . These aIn this perspective, we aim to explore possible sources of daily and circadian rhythmicity in the dopaminergic system. We first focus on abilities of dopaminergic neurons to express molecular and electrophysiological rhythms around 24 h. Then, we describe possible daily patterning of input to the dopaminergic system from other extra-SCN clocks. Finally, we aim to position the midbrain dopaminergic system as one of the hubs in the complex network of brain circadian oscillators.Per1, Per2, Per3, Bmal1 and Cry  and the number of spontaneously active dopaminergic neurons to change in a 24 h cycle (with the peak at the end of the dark phase) (Clock knockout) elevated both firing rate and bursting of dopaminergic neurons in vivo  . Additiok phase) . The actk phase) . Importak phase)  as well k phase) . The dis in vivo . OverallIn line with that, studies congruently demonstrate that dopamine concentration is elevated in the striatum during the active phase . HoweverClock-knockouts exhibit increased locomotor activity in the novel environment  serves as a relay between the SCN and the VTA . In retuAs mentioned above, the last 2 decades have brought discoveries of several extra-SCN neuronal populations of circadian oscillators with varying degrees of autonomy, which participate in shaping daily changes in physiology, behaviour, and cognitive processes. Unsurprisingly, at least some of these extra-SCN clocks are among the building blocks of the incredibly extensive, monosynaptic inputome of the ventral midbrain dopaminergic neurons . This maFirst, the circadian rhythmicity in the electrical activity present in these input structures may translate into changes of similar, hourly dynamics in the level of dopaminergic neuron firing and concomitant dopamine release in target brain regions as described above. Indeed, such circadian changes have been observed in multiple brain nuclei innervating dopaminergic neurons; with these neuronal centres being the source of those inputs spread throughout the whole cerebrum and carrying information of various kinds. Some of them are involved in maintaining homeostasis of the organism  changes in the basal activity of the dopaminergic system, as well as changes in the short-scale (occurring in milliseconds or seconds) phasic responses of DA neurons that encode value and/or salience of the perceived stimuli. Likewise, circadian changes in long-scale and short-scale dopaminergic activity should be apparent at the level of the animal’s behavioural states and behaviours.Importantly, daily changes in the dopaminergic system happening on the long timescale seem to depend predominantly on the intrinsic brain mechanisms ; notably, these are shaped by the long-lasting environmental conditions  . On the The pattern of electrical activity that a dopaminergic neuron exhibits at a given moment can be positioned on a spectrum ranging between two extremes: tonic, pacemaker-like firing of action potentials and, on the other end, generation of action potentials in purely bursting manner . These tOur current knowledge sheds light on the contribution of the circadian clock machinery to some of the slow changes observed in the activity of the dopaminergic system. Mostly, it describes the diurnal variation in the clock gene expression, overall electrical activity, concomitant dopamine release, as well as daily changes in the animal behaviour. These observations are consistent with the fact that in many neuronal populations circadian clock gene expression is accompanied, often causally, by changes in the expression of genes determining the membrane properties, neuron responsiveness to neurotransmitters, and the effectiveness of further signal transmission . In factAs already described above, dopaminergic system receives inputs from multiple regions distributed throughout the whole brain. Since presumably both the baseline activity and responsiveness to environmental cues of these inputs alter across day/night cycle, magnitude of dopaminergic system excitation/inhibition evoked by those inputs should be affected as well. More precisely, it is important to establish how the parameters  of responses of dopaminergic neurons to value/salience information provided by different inputs , ensures much more flexibility than a single, circadian input from the master clock. This assumption can be further strengthened by the fact, that these different “ticking” inputs are entrained by different environmental stimuli. With the SCN being a light-entrainable oscillator poorly entrained by feeding, the input from other extra-SCN clocks may provide information e.g., on the circadian patterning of food intake, valuable for the dopaminergic system functioning. Thus, considering the dopaminergic system as a hub in the intricate network of circadian oscillators, drawn in our perspective article, offers novel research opportunities in both reward and circadian neuroscience.


The diversity of cognitive task paradigms using functional near-infrared spectroscopy (fNIRS) and the lack of theoretical explanations for these functional imaging atlases have greatly hindered the application of fNIRS in psychiatry. The fNIRS brain imaging based on multiple cognitive tasks could generally reflect the working patterns and neurovascular coupling changes in the prefrontal working memory network. By alternating the stimulation patterns of resting and task states, six typical symptom-related functional brain imaging waveforms related to psychiatric disorders are identified and three joint networks of the prefrontal working memory, namely, the attentional working memory primary coordination network, the perceptual content working memory secondary network, and the emotional-behavioral working memory executive network, are initially represented. This is the first attempt to characterize the cognitive, emotional, and behavioral regulation of the prefrontal working memory network using fNIRS, which may promote the application of fNIRS in clinical settings. The improvement of diagnosis and treatment of psychiatric disorders relies on accessible neuroimaging tools that can identify associated frontal lobe dysfunction. Functional near-infrared spectroscopy (fNIRS) is an emerging functional neuroimaging method, non-invasively using the absorption and scattering relationships between multiple wavelengths of near-infrared light and chromophores in brain tissue  and decreased [deoxy-Hb], a process understood as part of the neurovascular coupling mechanism , 5. NeurAlthough NIRS can only observe the functional activity of about 2 cm of cerebral cortex under the probe, compared with PET, single-photon emission computed tomography (SPECT), fMRI, and other neurological effect technologies, it is relatively mobile, concise, and easy to meet measurement conditions, allowing subjects to be tested in a more natural and comfortable environment. As a result, more realistic data are obtained and could be used by psychiatrists. In recent years, several studies have shown that patients with psychiatric disorders have impaired function in the prefrontal cortex e.g., , 10]. Th. Th10)].The fNIRS, in conjunction with the verbal fluency task, holds promise for understanding, detecting, and differentiating psychiatric disorders, and has the potential for developing accessible neuroimaging biomarkers for different psychiatric disorders . The mulHowever, in China, it is generally found in clinical psychiatry that the current diagnostic criteria developed in Japan for unipolar depression and bipolar depression are of low diagnostic compliance in clinical application. Due to the low diagnostic compliance, the application of fNIRS in psychiatry is constrained. The low diagnostic compliance may be related to factors as follows: (1) there is only a single diagnostic reference imaging of bipolar and unipolar depression. It does not take into account the dynamic feature of the disease and the states of patients. With the remission of the disease, changes in brain functioning have an impact on the corresponding cerebral blood flow diagram. Some studies show that patients with depression show non-dominant hemisphere activation in thinking during depressive states and with their recovery from depression, a preference is returned to the dominant hemisphere in thinking . BesidesMulti-cognitive tasks include four cognitive tasks, namely the emotional picture identification task, (category) verbal fluency task, finger-tapping task, and setting-based sentence fluency task. The alternation of adaptive tasks and primary tasks as well as the alternation of resting and task states ensures the various representations of the prefrontal cortex in its cognitive regulation and cognitive dysfunction. A detailed introduction of tasks involved in the multi-cognitive tasks' paradigm is shown in During the multi-cognitive tasks, the relative concentration fluctuations of [oxy-Hb] and [deoxy-Hb] are recorded to reflect different mental disorders from a 45-channel fNIRS system  see .The symptoms (states) are identified and analyzed in the way of full-channel waveform, phase changes, and waveform composition ratios, and six typical waveforms see  with cliThe aim is to analyze functional abnormalities in the prefrontal cortex with fNIRS using various paradigms in psychiatry, with (letter) verbal fluency task as the primary task in the clinical diagnostic setting  and othOne of the primary functions of the prefrontal lobe is executive function. In empirical and theoretical papers, primary factors underlying executive functions are: (a) inhibition and switching –35, (b) In the neural network, the attentional working memory network is regarded as the main coordination network due to its importance. Attention is the taking possession by the mind, in clear and vivid form, of one out of what seem several simultaneously possible objects or trains of thought with the essence of focalization and concentration of consciousness. It implies withdrawal from some things in order to deal effectively with others  concentration. On the symptom classification mapping, the superposition of waveforms is seen, with an open hyperbola. The open positive phase indicates that the relative concentration of [oxy-Hb] at this site is higher than that of [deoxy-Hb]. The open negative phase indicates that the relative concentration of [deoxy-Hb] at this site is higher than that of [oxy-Hb]. Both of which may be a localized persistent pathological state. In adolescent children with attention deficit disorder or bipolar disorder with mixed episodes of inattention, and in mixed states of anxiety and depression, crossover disordered waveforms can be observed, i.e., the relative concentration curve of [deoxy-Hb] and the relative concentration curve of [oxy-Hb] are crossed, and the hyperbolic amplitude increases significantly increased and the waveforms are disordered.From the perspective of neural mechanism, almost all the current fNIRS research only analyzes the changes in the relative concentration of [oxy-Hb], and little is known about the effect of [deoxy-Hb] on the neurovascular coupling mechanism. Neurons, vascular smooth muscle cells/pericytes, and astrocytes are involved in neurovascular coupling. The destruction of physiological structures such as the peritubular space, blood–brain barrier, and humoral factors such as Ang can cause neurovascular coupling dysfunction. Part of the action mechanisms of different types of neurons, key neurotransmitters, receptors, and ions in related pathways are closely related to the maintenance of brain homeostasis because the neurovascular coupling mechanism widely exists in the brain and presents the characteristics of multi-structural participation and multi-channel regulation .It is generally believed that neurovascular coupling is only a marker of neuronal activation, with an amplitude proportional to the degree of neuronal activation, and changes in neurovascular coupling under disease are rarely considered. Csipo et al.  found thMultiple cognitive tasks analyze fNIRS imaging based on neurovascular coupling mechanism through cognitive tasks with different difficulties and divide the working memory network into three cognitive-related sub-networks for the first time. The near-infrared detection of multiple cognitive tasks presents the vascular coupling mechanism of the prefrontal neural network and neural working loops. Our articles –17 suppoThe subdivision of prefrontal working memory networks based on neurovascular coupling offers a novel insight into the analysis of fNIRS imaging representations, and also provides a direction for drug evaluation. This study may help expand the clinical application of fNIRS through the informed diagnosis of mental disorders and provide physicians with an objective measure that may eventually be used to personalize treatment. With the development of brain function imaging and imaging analysis technology, such as dynamic brain atlas, a more accurate and refined representation of the working memory network will be represented.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author.YR drafted the manuscript according to PL's clinical experience and was inspired by GC's psychological accounts. PL and YR discussed the theoretical framework and went through several minor changes. PL, GC, XZ, KF, and CY approved the final version of the manuscript with their guidance and comments on the article. 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.


Endophytes is a kind of microorganism resource with great potential medicinal value. The interactions between endophytes and host not only promote the growth and development of each other but also drive the biosynthesis of many new medicinal active substances. In this review, we summarized recent reports related to the interactions between endophytes and hosts, mainly regarding the research progress of endophytes affecting the growth and development of host plants, physiological stress and the synthesis of new compounds. Then, we also discussed the positive effects of multiomics analysis on the interactions between endophytes and their hosts, as well as the application and development prospects of metabolites synthesized by symbiotic interactions. This review may provide a reference for the further development and utilization of endophytes and the study of their interactions with their hosts. Endophytes are special “microorganism” with much value. They can parasitize different parts of living plants but do not obviously cause symptoms of external infection in host plants or symbiosis and coevolution with their host . As an iExploring the application of endophytes has become a hot topic with the gradual understanding of the interaction between endophytes and hosts. Plants were first thought of as individual organisms that produced metabolites by themselves to completely support their growth and development. This cannot explain the phenomenon in which plants growing in the wild are “better,” such as in medicinal characters, than those under artificial cultivation; artificial cultivation of medicinal plants loses the original effect, etc. It was not until the discovery of endophytes that their interactive relationship was gradually revealed, especially after the application of omics technology at the cellular and molecular levels in which the study of the interactive relationship between the two progressed rapidly. In recent years, researchers have been trying to explore the evolutionary relationship between host and endophytes, and trace the reciprocity mechanism of this specific evolutionary relationship, in order to provide theoretical basis for the commercial development of endophytes. At present, some endophytic bacteria and their active products have been successfully used in commercial production and obtained great benefits in the preparation of new drugs and agricultural protection . It is oTo provide a reference for revealing the complex interaction between endophytes and their hosts, this review first discusses the interaction between endophytes and host plants and then the effects of endophytes and their secondary metabolites on plant growth and development. The application of endophyte resources instead of is also discussed.Many plant biological functions are dependent on endophytes, and each plant is actually a complex microecosm . A broadCamptotheca acuminata kills invading microorganisms by producing camptothecin, which competitively inhibits the activity of the microbial topoisomerase I complex , peptidoglycan (PG), superoxide dismutase (SOD), β-chitosan bacteria cold shock protein (RNP1motif), chitin, etc.] and the specificity of the host cell surface receptor molecules PRR identification  production in the host. ISR differs from traditional system acquired resistance (SAR) in that its phenotype is similar to that of pathogen induced SAR, both of which can induce broad-spectrum resistance to pathogens in plants . In essefication . Under cfication . In factfication . After pfication . Howeverfication . Existinfication .P. indica as a plant probiotic preparation. Genome-wide analysis of the endophytic bacterium Pantoea ananatis revealed the existence of genes encoding hydrolase, n-acyl high serine lactone synthase, superadhesion factor and fusylic acid resistance protein. This shows its great biological potential for commercial production , receptor-like kinases (RLKs), ion channels, and histidine kinases, when the host is exposed to salinity, temperature, drought, and heavy metal toxicity. Change the concentration of Ca2+ in the cytoplasm and produce corresponding signaling molecules such as reactive oxygen species (ROS), ABA and inositol phosphates (IPs). Further kinase responses activate various downstream transcription factors, such as MYC/MYB, WRKY, DREB/CBF, AREB/ABF, bHLH, NAC, and bZIP (LTPs), heat shock proteins (HSPs), late embryogenesis abundant protein (LEA), antioxidant response element (ARE) and osmotic proteins, through reciprocal transformation of protein phosphorylation and dephosphorylation. Some endophytes activate pattern recognition receptor (PRR) membrane receptors under the influence of exogenous signals, amplify secondary signals through cascade reactions, and then activate the downstream MAPK signaling pathway, resulting in NLRs, TFs, HSFA2, RLKs and other related gene expression. Through the above series of biochemical reactions, endophytes can regulate host permeability to reduce or even offset the effects of stress and can finally achieve the purpose of repairing damage , which can induce systemic resistance and antibiotics, inhibit rice bacterial diseases and promote rice growth and development. The endophytic bacterium Bacillus subtilis EDR4 isolated from wheat competitively inhibited the growth of pathogens Fusarium oxysporum by producing the antifungal protein E2 -like superfamily proteins, Lateral organ boundaries (LOB) domain proteins and Broad-complex (BTB)/pox virus (POZ)/PDZ-binding motif (TAZ) domain proteins may play a role in the growth promotion of wild Chinese sweet grass . The study of the mechanisms of interactions between endophytes and their hosts, such as the specific ways of endophyte colonization, whether the endophyte can achieve proliferation after colonization and whether their host has antagonistic reactions, is still unclear. In terms of host growth and development, how endophytes regulate host metabolism, such as producing new metabolites and “reprocessing” host metabolites, still needs to be further revealed. In terms of production and application, problems such as difficulty in breeding on a large scale in vivo from their host. In vitro endophyte isolation, culture and even fermentation cannot accurately obtain the corresponding strains or even metabolites. In addition, there are interactions between endophytes in the host, which undoubtedly adds difficulties in endophytes study. The mystery of endophytes is gradually being revealed. In future, on the basis of ensuring the biological activity of isolated and cultured endophytes, the improvement of their characteristics and application in basic research and commercial production will be of great significance. For example, new drugs can be created to treat diseases and for agricultural production. Regulating host gene expression and pathways improves the growth of valuable medicinal plants, realizing transgenic breeding and improving crop quality. Now is the time to elevate endophyte research from traditional physiological and biochemical research to higher cellular- and molecular-level research. Combined with omics technology, a database of endophytes and their active metabolites should be established. Then, the database should be used to understand the unknown field of endophytes and host interactions and to benefit from it.Solving the above problems will not only help to tap into the ecological functions of endophytes but will also further improve the application potential of endophytes and will provide a base for the further development and utilization of endophytes. At present, although there have been an increasing number of studies on endophytes, there have been few reports on the large-scale application of endophytic preparations and their active metabolites in commercial production. The utilization of endophyte biological resources is still difficult, and there is no effective detection technology to directly identify endophytic bacteria DX and LL conceived, supervised, and wrote and reviewed the manuscript. ZL, WW, YH, and MQ originally wrote and reviewed the draft. LL, DX, and YH co-founded and co-administrated the project. All authors read and approved the final 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.