Aging is considered a risk factor for various diseases including cancers. In this aging society, there is an urgent need to clarify the molecular mechanisms involved in aging. Wnt signaling has been shown to play a crucial role in the maintenance and differentiation of tissue stem cells, and intensive studies have elucidated its pivotal role in the aging of neural and muscle stem cells. However, until recently, such studies on the gastrointestinal tract have been limited. In this review, we discuss recent advances in the study of the role of Wnt signaling in the aging of the gastrointestinal tract and aging-related carcinogenesis. Aging is one of the most intriguing themes of research in the field of life science, especially in this aging society, since it causes deterioration of organic functions and leads to various diseases, including life-threatening cancers . UnderstWnt signaling is a major signaling pathway that has been intensively studied ,10,11. TIn the absence of the Wnt ligand, cytosolic β-catenin binds to the β-catenin destruction complex, which includes the scaffolding protein Axin, adenomatous polyposis coli (APC), serine/threonine kinase glycogen synthesis kinase 3 (GSK3), and casein kinase 1 (CK1). The β-catenin is then phosphorylated and becomes ubiquitinated by E3-ubiquitin ligase β-transducin repeat containing protein (β-TrCP), and, finally, is subjected to proteasomal degradation . On the In the gastrointestinal tract, Wnt and other Wnt-related factors are considered to be secreted mainly from mesenchymal cells located in proximity to gastrointestinal stem cells . In respThe role of this canonical Wnt signaling in the development of cancer has been intensively studied ,16. PrevOne of the main causes of aging is the aging of tissue stem cells . In the Cho and colleagues have shown that inhibition of sFRP3, a Wnt antagonist previously reported to accumulate and suppress quiescent neural stem cells in the adult brain , counterWith respect to studies of stem cell aging in skeletal muscle, Brack and colleagues reported that muscle stem cells of aged mice tended to convert to fibrogenic lineages as a result of enhanced canonical Wnt signaling and that this tendency could be suppressed by Wnt inhibitors . SimilarHair follicles are another tissue in which stem cells have been vigorously investigated. Castilho and colleagues showed that, although activation of Wnt signaling induced growth of hair follicles, prolonged exposure to the Wnt ligand led to exhaustion of stem cells and hair loss, suggesting that enhanced Wnt signaling induced stem cell aging in hair follicular stem cells .The preceding studies have demonstrated that suppressed or enhanced canonical Wnt signaling regulates stem cell aging in neural, muscular, and hair tissues. Similarly, it has become evident that Wnt signaling plays an important role in gastrointestinal stem-cell aging which we discuss below.The studies described have shown that Wnt signaling plays a crucial role in regulating the aging of neural, muscular, and hair follicular stem cells. Next, we consider the role of Wnt signaling in regulating gastrointestinal stem cells.It is now widely accepted that leucine-rich orphan G-protein coupled receptor 5 (Lgr5) is a marker for intestinal stem cells residing at the base of intestinal crypts . Lgr5 waLgr5 is a G-protein-coupled receptor that serves as a receptor for R-spondins . R-spondStem cell markers in the stomach have been reported to differ between the antrum and the corpus. In the antrum, Lgr5, Axin2, and Aquaporin5 (AQP5) have been reported as markers for gastric stem cells located in the base of antral glands ,33,34, wHence, although the stem cell marker for gastric stem cells is still controversial , these eSo, how does Wnt signaling affect aging of the gastrointestinal tract? A study performed by Nalapareddy and colleagues investigated the influence of aging on intestinal stem cells . They shNotum, as well as Wnt3a supplementation, restored the function of aged intestinal organoids, and pharmacological inhibition of Notum also restored the regenerative capacity of the aged murine intestine. Hence, this study also indicated that decreased Wnt signaling in intestinal stem cells was the underlying factor in the aging of the intestine.Another recent study from Pentinmikko and colleagues on Wnt signaling and intestinal stem cells also reported that Wnt signaling was reduced in aged intestinal stem cells . The autOn the other hand, Cui and colleagues reported the impact of augmented Wnt signaling on aged intestinal stem cells . They shThe studies above, although through a different mechanism, proposed that Wnt signaling in intestinal stem cells plays an important role in the aging of the intestinal tract. In particular, decreased Wnt signaling in aged intestinal stem cells led to impaired regenerative capacity. However, there are other reports which suggest correlation of enhanced Wnt signaling and aging of the gastrointestinal tract.One of these studies was performed by Hayashi and colleagues, who investigated the effect of aging on the motility of the stomach. They found that the number of interstitial cells of Cajal (ICC) was decreased in the stomachs of aged mice and that this decrease was associated with impaired gastric compliance . It was ARF, p53, and p21WAF1 (Dkk3 gene. Hence, consistent with the report from Hayashi and colleagues, we observed correlation of enhanced Wnt signaling and aging of the gastrointestinal tract.Recently, we reported further evidence of a correlation between enhanced Wnt signaling and aging in the stomach . We foun p21WAF1 . FurtherTaken together, the studies described above indicate that Wnt signaling plays an important role in the aging of the intestine and the stomach.Aging is considered one of the risk factors for cancers in various organs . RegardiTCF4, a transcription factor that functions as the downstream effector of canonical Wnt/β-catenin signaling in normal gastric mucosa was reported to increase with age [TCF4 enhanced proliferation of cells in in vitro studies using a gastric cancer cell line, showing that Wnt signaling altered proliferation in aged gastric mucosa.Epigenetic reprogramming in stem cells has also been reported to induce malignant neoplasm in aging stem cells , and metwith age . The repDkk3 gene in aged gastric organoids, the cells originating from the aged gastric mucosa, led to enhanced Wnt signaling and Tbx3 induction, resulting in increased proliferation due to suppressed cellular senescence [In our recent study, we showed that DNA methylation of the nescence . Regardinescence . They foThe studies discussed above strongly indicate that the Wnt signaling pathway is involved in the aging of the gastrointestinal tract. In addition, our study has suggested a possible mechanism contributing to aging-related carcinogenesis in the stomach, namely, the Dkk3-Wnt-Tbx3 pathway . Since tTranscription factor TBX3 was first identified as the responsible gene for ulnar-mammary syndrome, a rare pleiotropic disorder affecting limb, apocrine gland, tooth, and genital development . It funcOther Wnt signaling inhibitors have also been considered as therapeutic reagents for solid tumors, including gastric cancers and colorectal cancers, and hematological malignancies ,65. WNT9As discussed in this review, it is evident that Wnt signaling plays a crucial role in the aging of the gastrointestinal tract. However, whether enhanced or diminished Wnt signaling is related to aging is still controversial. It is possible that it depends on the organs involved, such as observed in Wnt-suppressed aged neural stem cells and Wnt-enhanced aged muscular stem cells, but further studies are awaited to elucidate this issue.


Although productive progress has been made in colorectal cancer (CRC) researchs, CRC is the second most frequent type of malignancy and the major cause of cancer-related death among gastrointestinal cancers. As angiogenesis constitutes an important point in the control of CRC progression and metastasis, understanding the key signaling pathways that regulate CRC angiogenesis is critical in elucidating ways to inhibit CRC. Herein, we comprehensively summarized the angiogenesis-related pathways of CRC, including vascular endothelial growth factor (VEGF), nuclear factor-kappa B (NF-κB), Janus kinase (JAK)/signal transducer and activator of transcription (STAT), Wingless and int-1 (Wnt), and Notch signaling pathways. We divided the factors influencing the specific pathway into promoters and inhibitors. Among these, some drugs or natural compounds that have antiangiogenic effects were emphasized. Furthermore, the interactions of these pathways in angiogenesis were discussed. The current review provides a comprehensive overview of the key signaling pathways that are involved in the angiogenesis of CRC and contributes to the new anti-angiogenic strategies for CRC. Colorectal cancer is the second most frequent type of malignancy and the major cause of cancer-related death among gastrointestinal cancers, despite significant advances over the past two decades in preventive screening and therapy aimed at improving patient survival . The oveJudah Folkman’s early observations showed that fast-growing tumors have a large number of blood vessels compared to dormant tumors, leading to the concept that tumor progression requires the initiation of tumor angiogenesis and anti-angiogenesis . ThroughAs a hallmark of cancer, angiogenesis is closely related to tumor growth, metastasis, invasion, prognosis, and recurrence ,6,7,8. TAngiogenesis is a crucial process in CRC development, in which VEGF signaling pathway is a classical regulatory pathway in CRC . Thus, iThe VEGF signaling pathway is established as one of the key regulators of tumor angiogenesis . The VEGMany biologically active molecules promote angiogenesis by regulating the VEGF signaling pathway. It has been well established that the HIF-1 was a key mediator of hypoxic responses. Under hypoxia, HIF-1 could significantly upregulate the expression of VEGF ,21. As tIn addition, some molecules that inhibit angiogenesis through the VEGF signaling pathway have been described. These include chemokine CC ligand 19 (CCL19), also known as macrophage inflammatory protein 3-β (MIP-3B). The expression of CCL19 in CRC tissues was lower than that in normal tissues, and negatively correlated with cancer angiogenesis. CCL19 inhibited the Met/ERK/ELK-1/HIF-1α/VEGFA pathway by promoting the expression of miR-206 . CCL19 iErycibe obtusifolia Benth, inhibited angiogenesis by blocking the autophosphorylation and downstream signaling pathways of VEGFR2 . As. As74]. In addition, some inhibitors of NF-κB have a similar effect of suppressing angiogenesis in CRC. PDTC could inhibit CXCL5-dependent induction of FOXD1 and VEGFA expression . BAY11-7It has been reported that imatinib could activate different intracellular signaling pathways, thereby breaking the feedback loop between proinflammatory cytokines and transcription factors . Imatinib also appeared to inhibit the coordination of proinflammatory cytokines by intracellular signaling, which was also involved in the upregulation of angiogenic factors in CRC  48]. As. As48]. Prevention of angiogenesis by NF-κB-specific inhibitors has been the core of anticancer therapy. Andrographolide (AP), a natural phytochemical found in Andrographis paniculata, antagonized IL-8 induced by TNF-α by inhibiting NADPH oxidase/ROS/NF-κB and other signaling pathways, which led to the inhibition of angiogenesis in the tumor microenvironment  51]. Pa. Pa51]. The JAK/STAT signaling pathway plays a critical role in various aspects of CRC, especially angiogenesis in CRC. This pathway consists of tyrosine kinase-associated receptors, JAK and STAT, which are coupled to activate. By binding with tyrosine kinase-associated receptors, over 40 different cytokines or growth factors can induce the STAT3 signaling pathway. The JAK family proteins include four members, JAK1, JAK2, JAK3, and TYK2, related to the cytoplasmic regions of tyrosine kinase-associated receptors . STAT faThe end of the active STAT signaling pathway enables a succession of gene expression changes, which then participate in biological processes, including proliferation, angiogenesis, and metastasis ,86,87,88Promoting factors in the JAK/STAT signaling pathway play a very important role in the regulation of CRC angiogenesis. These biologically active molecules include cytokines (chemokines), various proteins, and lncRNAs . The agoSolute carrier family 6 member 14 (SLC6A14) expression, which is low in normal human cells, is subject to the change of the JAK2/STAT3 pathway in CRC ,98. SLC6In terms of the lncRNA FLANC, there are connections with the JAK/STAT3 signaling pathway. FLANC is a primate-specific lncRNA residing within the first intron of Cadherin EGF LAG seven-pass G-type receptors 1 (CELSR1), which is weakly expressed in normal colon cells . This geVarious factors inhibiting angiogenesis via the JAK/STAT3 signaling pathway in CRC have been described. These include proteins, plant-based compounds, and RNAs .+/H+ exchanger regulatory factor 2 (NHERF2) is an angiogenesis-inhibiting protein. The NHERF family of proteins are scaffolds that orchestrate the interaction of receptors and cellular proteins . C. C132]. c-Myc [c-Myc, and cyclin D1, which inhibited the angiogenesis of CRC [Dickkopf-1 (DKK-1) is a secreted protein and an extracellular inhibitor of the Wnt signaling pathway . DKK-1 bc-Myc . LncRNA s of CRC .Evidence suggests that the synthetic role of miR-29b was essential for the inhibition of CRC cells . MiR-29b2D3) has the potential as a therapy for CRC by inhibiting the angiogenesis through the Wnt pathway  in the full length of the colon crypts [Vitamin D  pathway . Vitamin pathway . With thn crypts .B-cell CLL/lymphoma 9 (BCL9) was regarded as a co-activator of the Wnt/β-catenin signaling pathway by participating in TCF-mediated transcription in CRC . StabiliThe Notch signaling pathway is one of the most important signaling modes that plays a part in physiological and tumor pathology. It is not only essential for differentiation, proliferation, and apoptosis, but also angiogenesis, tip/stalk cell selection, and arteriovenous specification ,145. TheThe Notch signaling pathway is regularly altered in many cancers, as well as CRC . In the It is highly important to explore the factors that inhibit angiogenesis in CRC by inhibiting the Notch signaling pathway. γ-Secretase and Dll4 are the most common targets of small-molecule inhibitors or antibodies for the Notch signaling pathway blockade . An incrTargeting the Notch signaling pathway to prevent angiogenesis in CRC is feasible . As a naMultiple pathways commonly interact during angiogenesis in CRC. Current studies found that the Notch signaling pathway was associated with the Wnt and VEGF signaling pathways in CRC angiogenesis. Many studies have suggested that the Wnt/β-catenin/TCF signaling pathway positively regulated Jagged1 expression to activate Notch expression in CRC . As an aSeveral studies have confirmed that NF-κB could interact with VEGF to jointly promote angiogenesis. These interactions included the aforementioned B7-H3/NF-κB/ VEGFA axis and the AKT/NF-κB/FOXD1/VEGFA pathway ,69. FurtIn addition to the above, some compounds could influence interactions in multiple pathways. Enalapril could significantly enhance the sensitivity of CRC to 5-FU and its antitumor effect by inhibiting NF-κB/STAT3 regulatory protein, proliferation and angiogenesis . Crocin In this review, we summarized the pathways that influence angiogenesis in CRC. The specific content includes the basic introduction of the pathway, mechanism, some promoting and inhibiting factors of the pathway, targeted therapy and crosstalk. This review introduced not only the interactions among signaling pathways and some drugs already in clinical use but also the natural compounds that affect the crosstalk of multiple signaling pathways, which have great value in the research of targeted therapy for signaling pathways. We anticipated that these natural compounds could lead to new directions in the treatment of CRC in the future. Although many studies have focused on the role of signaling pathways in CRC angiogenesis, further research is needed to find more effective therapeutic agents.


Transgender people are just as able to be good parents as anyone else. The treatment involved in addressing their gender dysphoria usually removes their natural fertility and if they want to preserve their fertility, they will need gamete storage. The technology needed to provide gamete storage for transfolk is not any different from anyone else but the clinic setup, human interactions and the approach used need to be sensitive and require everyone in the clinic to understand the issues and behave accordingly if high-quality service is to be provided. Transpeople are individuals who are not content to live in the gender role that follows from the sex they were assigned on their birth certificate. Instead, they make a change to a different social gender role – usually, the one not assigned on their birth certificate but sometimes, more rarely, to a gender-neutral role. These individuals attract an International Classification of Diseases version 11 diagnosis of gender incongruence .with, the other who you go to bed as.Being trans is not a mental illness or disorder – gender incongruence is ‘a condition related to sexual health’ according to the International Classification of Diseases version 11. There is no connection to sexual orientation – one is who you go to bed Nearly all transpeople who have made a change of social gender role seek endocrine treatment. Transwomen take estradiol to elicit an estrogen level in the female follicular range and very often a gonadotrophin-releasing hormone analog to suppress testicular testosterone levels into the normal female range. Transmen take testosterone to elicit a testosterone level in the normal male range, which generally serves to suppress ovarian function. Non-binary people generally seek a hormonal profile somewhere between these two states.Nearly all transmen undergo a bilateral mastectomy and male chest reconstruction but only about a third seek the complicated, multi-stage surgery needed to create a penis  which often incorporates a hysterectomy and bilateral salpingo-oophorectomy.About two-thirds of transwomen seek genital reconstructive surgery – which involves the creation of a vulva and clitoris and very often a neovagina as well. Many self-fund augmentation mammoplasties or facial feminization surgery as neither procedure is currently NHS funded.The mean age at presentation to a gender identity clinic has fallen over the years and is now such that, for the majority of patients, fertility preservation is an issue. A growing proportion is so young as not to have thought seriously about family formation, always having felt that at their age it is ‘in the future’, but suddenly finding themselves having to think about it immediately, as their natural fertility is threatened by hormone treatment. Impatience to start hormone treatment, with later regret about lost fertility or the need to interrupt treatment to preserve fertility, is a fairly common problem.Most transpeople seeking to preserve their fertility want to do so before commencing hormone treatment. A few suspend already well-established hormone treatment to preserve their fertility, sometimes having changed their minds late in the day or having been earlier wrongly advised that NHS funding was not available. A very small number suspend completed hormone treatment to impregnate a partner or to themselves become pregnant.Getting it right for transpeople starts long before the patient actually arrives in your clinic. Has the prior communication used the appropriate pronoun and name?  Do not send a transman pre-appointment information leaflet that begins ‘when a woman has her eggs collected she...’ but instead consider an across-the-board mild and completely grammatically sound conversion so instead it reads ‘when people have their eggs collected they...’ This re-wording works for every patient having eggs collected, transman and woman alike. Note also that the Human Fertilisation and Embryology Authority provides forms, especially for use by transpeople, that you must use  needing to use those same facilities. Ask the women in your department whether the facilities currently used to collect semen samples are a place where they would be comfortable, as a woman, masturbating to orgasm. If they are not, you need to change them until they are. Note also that while most transwomen can produce a semen sample by masturbation, a significant proportion, particularly younger patients, simply cannot do this in any setting, including the privacy of their own home. These patients may need surgical sperm retrieval or electroejaculation, as used for peripubertal boys with childhood malignancies.Avoid genital exposure or examination unless it is absolutely necessary and there is no alternative. Many transpeople cannot bear to look at their own genitals, let alone let anyone else do so. Always use abdominal scans if it is in any way possible and if the genital examination is unavoidable, consider sedation or anesthesia. If the patient is conscious, bear in mind that many transmen would much prefer an ultrasound probe placed rectally rather than vaginally. Some would rather prefer a man wielding that alarming-looking probe, some a woman. As ever, the golden rule is to ask, to offer a choice if at all possible and then to go with that choice.In a related vein, for transmen, the area between the shoulder sockets and above the xiphisternum is ‘the chest’ whether or not there are what that look to you like breasts on it. What looks like ordinary breasts to you are unwanted by him and feel alien and inappropriate. Use the term he feels comfortable with, not one that reminds him of his unwanted problems.Notably, fertility preservation in the context of gender incongruence is funded throughout Scotland and Wales and by virtually all English Clinical Commissioning Groups (CCG). In April 2019, NHS England advised Clinical Commissioning Groups that ‘all patient groups whose medical treatment may compromise fertility should be in the contemplation of a CCG when its clinical commissioning policy for fertility preservation is being developed or is under review. Given the legal duties identified above, CCGs must not determine which patient groups might be offered fertility preservation services on a basis that discriminates against those patients because of a protected characteristic, including gender re-assignment’ . Because their gametes were preserved, they are to be used for inter-partner donation and the patients must be advised that because they are acting as a donor, the laws stipulated by the HFEA will oblige them to undergo the relevant screening processes.The author declares that there is no conflict of interest that could be perceived as prejudicing the impartiality of this commentary.This study did not receive any specific grant from any funding agency in the public, commercial or not-for-profit sector.


On 31 May 2021, the Political Bureau of the Central Committee of the Communist Party of China proposed the policy that a couple can have three children, and rolled out more supportive measures to further optimize the fertility policies. However, while the Chinese government is further optimizing its fertility policy, the sudden outbreak of COVID-19 is raging around the world, which threatens the implementation of China’s fertility optimization policy. Based on this, this paper firstly explores the impact of COVID-19 on women’s fertility intentions. Secondly, based on the Theory of Planned Behavior, this paper constructs a structural equation model to quantitatively reveal the specific factors that affect women’s fertility intentions under the epidemic, as well as their impact paths, and then puts forward corresponding suggestions for the government to solve the problem of fertility, aiming at delaying population aging and optimizing population structure. The research results show that: (1) COVID-19 lowers the fertility intentions of women of childbearing age. (2) During the pandemic, economic pressure emerged as the biggest factor affecting women’s fertility intentions. The decline in income caused by the pandemic has become an important factor in preventing women from having children. (3) The conflict between work and childbearing is still an important factor affecting the fertility intentions of women of childbearing age. The government’s provision of perfect childcare services and their strengthening of the protection of women’s employment rights and interests will greatly reduce women’s anxiety about childbearing. Since the 1980s, the world’s major developed countries have generally experienced reduced fertility rates, even falling below the replacement level, and long-term fertility sluggishness . At presThere are many studies on the impact of COVID-19 on fertility intentions, but most existing studies have focused on European and American countries affected by more severe epidemics. For example, Naya et al.  found thBased on this, this paper constructs a structural equation model to comprehensively analyze the influences of various factors on the fertility intention of Chinese women during the epidemic period, and further proposes specific suggestions for the government to optimize fertility policies and implement fertility support measures to actively respond to the trend of population aging and expand the scale of fertility.The paper is organized as follows: Fertility intention is a significant factor affecting the actual fertility level of families and the total population. First, it is clear that although there is no unified standard for the concept of fertility intention and the quantification of its indicators, scholars involved in the process of discussing fertility intentions have recognized that it is not a static desire, but a tendency of individuals to have children in real life, which is constrained by real conditions in terms of the number of children, the timing of childbirth, the gender, and even the healthiness of children. For example, Wu  argued tMany scholars have fully studied the changes of fertility intention during the epidemic. For instance, Druetz et al.  verifiedIn summary, there has been a great deal of research on fertility intentions conducted by both domestic and international scholars, focusing on the concept of fertility intentions and the analysis of influencing factors. However, most studies have only explored the impact of a single factor on fertility intention during the pandemic, and few studies have comprehensively considered the impacts of various factors on fertility intention. Based on this, this paper selects women of childbearing age as the survey objects, uses questionnaire data to analyze the changes in women’s fertility willingness during the epidemic, builds a structural equation model to comprehensively analyze the influence of various factors on fertility intentions during COVID-19, explores the impact of the epidemic on their fertility willingness, and provides further information for the government. Suggestions are made to improve supporting measures for fertility, so as to meet the real needs of women in terms of fertility and increase their willingness to bear children.The questionnaire is mainly composed of three parts: deriving the basic information of the respondents, a survey on the current situation of fertility intentions, and an analysis of factors affecting fertility intentions during the epidemic.First, referring to the research of Zhuang , the demSecond, in the survey on the current situation of fertility intentions, this paper investigates the current fertility intentions of the respondents in two aspects: the expected number of children and the short-term fertility intentions. Molica et al.  conducteThird, to quantitatively reveal the specific factors affecting women’s fertility intentions during the epidemic, the scale is divided into eight dimensions, namely, economic support, public support, policy support, family support, perceived risk, perceived behavioral control, behavioral attitudes, and fertility intentions.A specific questionnaire can be seen in the A total of 443 valid questionnaires were collected online between 7 January 2022 and 21 April 2022. During the period, the more infectious COVID-19 variant strain Omicron showed an outbreak trend in Beijing, Tianjin, Shanghai, Hong Kong, Jilin, and Shaanxi, with a cumulative total of 589,870 confirmed cases , and a cumulative total of more than 890,000 people were centrally quarantined in Shanghai due to the outbreak (source: The Paper). After the release of the questionnaire, COVID-19 did not stop spreading, and appeared in Beijing and Shanghai, with 710,000 people stuck in the Shanghai containment area . Based on this, the fertility intentions of women surveyed by the questionnaire released during this period will be interpreted more diversely. Through the online platform, we collected questionnaires from all over the country. However, the number of women of childbearing age (between 22 and 46 years old) in China is about 240 million (data source: China’s sixth Census in 2010), and the sample size of this paper is 443, so there is some bias in the sample selection.After the descriptive statistics of the valid questionnaires, the specific distribution of the sample is shown in Regarding the fertility intentions of the survey respondents, this section focuses on two perspectives: the number of expected children and short-term fertility intentions.The number of expected children as been derived via the questionnaires, and the results are shown in The results of the short-term fertility intentions of women of optimal age to conceive a child are shown in The impact of COVID-19 on women’s fertility intentions is studied in this paper. Currently, the change in the fertility intentions of the female population due to the epidemic is the key variable in this questionnaire, and the variable interpretation plot is shown in As shown in The results of the above descriptive statistical analysis show that the epidemic has negatively impacted the fertility intentions of women of childbearing age to some extent. The next section will further explore the specific factors affecting the fertility intentions of women of childbearing age, and their influence paths as affected by the epidemic.This section proposes research hypotheses based on the Theory of Planned Behavior and related studies, and constructs a theoretical model. In addition, it analyzes the reliability of the scale questionnaire using the SPSS26.0 software , tests the convergent validity and differential validity of the scale questionnaire using AMOS24.0 software , conducts a descriptive statistical analysis of the factors, and constructs a structural equation model of the factors affecting women’s fertility intentions. This section reveals the factors influencing women’s fertility intentions from a quantitative perspective and analyzes their influence paths.In 1985, Ajzen added perceived behavioral control variables to the original rational behavior framework, thus forming the Theory of Planned Behavior (TPB).The TPB aims to explain human behavior rather than just predict it, and it considers that behavioral attitudes, subjective norms, and perceived behavioral control are the three prerequisites for determining behavioral intentions and behavior. The behavioral attitude refers to an individual’s positive or negative value judgment of behavior. The subjective norm refers to the social pressure individuals feel due to a particular behavior. The perceived behavioral control refers to an individual’s perception of the difficulty level of an operating behavior. Based on this, the subjective attitudes toward fertility, perceptions of family and friends’ attitudes, external social norms, and cost constraints determine the behavioral attitudes, normative beliefs, and perceived behavioral control of fertility.Since its initial introduction, the Theory of Planned Behavior has been widely used in various industries, such as healthcare, leisure activities, travel, environmental behaviors, employment choices, shopping consumption, online activities, and online services. The predictive and explanatory power of the three variables in the model, behavior attitudes, subjective norms and perceived behavioral control, in relation to behavioral intentions has been empirically demonstrated in several studies. Ajzen et al.  analyzedHypothesis 1 (H1).Perceived behavioral control has a significant positive effect on fertility intentions.Hypothesis 2 (H2).Behavioral attitude has a significant positive effect on fertility intentions.For the selection of external variables, this paper selected economic support, policy support, public support, family support and perceived risk as external variables for the following reasons.Therefore, research hypotheses H3-H11 are proposed, and a theoretical model of the fertility intentions of women of childbearing age is constructed, as shown in Hypothesis 3 (H3).Economic support has a significant positive effect on perceived behavioral control.Hypothesis 4 (H4).Perceived risk has a significant negative effect on perceived behavioral control.Hypothesis 5 (H5).Family support has a significant positive effect on perceived behavioral control.Hypothesis 6 (H6).Policy support has a significant positive effect on perceived behavioral control.Hypothesis 7 (H7).Public support has a significant positive effect on perceived behavioral control.Hypothesis 8 (H8).Economic support has a significant positive effect on behavioral attitudes.Hypothesis 9 (H9).Family support has a significant positive effect on behavioral attitudes.Hypothesis 10 (H10).Policy support has a significant positive effect on behavioral attitudes.Hypothesis 11 (H11).Public support has a significant positive effect on behavioral attitudes.In this section, specific questions are designed for the eight sub-active variables of the questionnaire, and the latent variables and their codes are shown in (1) Reliability analysisThe reliability analysis, also called the reliability test, measures and tests the reliability and stability of the questionnaire. Cronbach’s alpha (α), which is commonly used in academia, is adopted to analyze the intrinsic reliability of the scale. Generally speaking, the closer the alpha value is to 1, the better the reliability of the questionnaire. When α is greater than 0.8, the reliability of the questionnaire is high. When α is 0.7–0.8, the reliability of the questionnaire is good. When α is 0.6–0.7, the reliability of the questionnaire is acceptable. When α is less than 0.6, some problems exist in the design or structure of the questionnaire, which should be revised.The results of the reliability test of each variable in the questionnaire, performed by SPSS 26.0, are shown in (2) Validity analysis(1) Convergence validity analysisConvergence validity analysis is carried out by analyzing the factor loadings of each dimension. In this paper, the convergence validity of the questionnaire is determined by three indicators: standardized loading coefficient (Std.), average variance extracted (AVE), and combined reliability (CR). The questionnaire scale has convergence validity when Std. > 0.5, AVE > 0.5 and CR > 0.7. As can be seen from (2) Distinguishing validity analysisDistinguishing validity analysis is used to estimate the difference in the degree of correlation between a measure and different internal and external structural variables by comparing the internal correlation of the measure within a structural variable with the external correlation of each structural variable, and thus determining the distinguishing validity of the variable. If the square root of the AVE of a variable is greater than the correlation coefficient of that variable with other variables, then the distinguishing validity of that variable is good. As can be seen from The descriptive statistical analyses for each factor score are shown in In summary, the fertility intentions of women of childbearing age during the epidemic are influenced by several factors, and the specific influence paths of each factor affecting fertility intentions will be explored in the next section using structural equation modeling.(1) Evaluation of the overall fitness of the structural equation modelStructural equation modeling (SEM) is an empirical analysis technique used to find the relationship between variables and verify whether the theoretical model and hypotheses are reasonable. Based on the research hypothesis and the theoretical model constructed in the previous paper, AMOS24.0 software is used to construct a structural equation model of the factors influencing women’s fertility intentions. The relative chi-square (CMIN/DF), comparative fit index (CFI), Tucker–Lewis Index (TLI), and root mean square of the error of approximation (RMSEA) are used as fitted indicators to test the fitness of the structural model. As can be seen from (2) Test results of the research hypothesisThe results of the structural equation model of “factors influencing women’s fertility intentions”, the model relationships and standardized path coefficient estimates among the latent variables, as well as the hypothesis tests, are shown in p values of perceived behavioral control and behavioral attitude related to fertility intentions are less than 0.001, and the standardized estimates are positive, indicating that perceived behavioral control and behavioral attitude have a significant positive effect on fertility intentions, so the original hypotheses H1 and H2 are accepted. The standardized path coefficient of behavioral attitude affecting fertility intentions is 0.611, which is significantly greater than the standardized path coefficient of perceived behavioral control (0.236), indicating that behavioral attitudes have a greater effect on fertility intentions.As shown in p values of economic support and public support in relation to perceived behavioral control are 0.006 and 0.007, respectively, both of which are less than 0.05, and the standardized estimates are greater than 0, indicating that economic support and public support have a significant positive effect on perceived behavioral control, so the original hypotheses H3 and H7 are accepted. The p values of perceived risk related to perceived behavioral control are less than 0.001 and the standardized estimates are less than 0, indicating that perceived risk has a significant negative effect on perceived behavior control, so the original hypothesis H4 is accepted. Hypothesis H5 is not tested, indicating family support has no significant effect on perceived behavioral control, because most of the respondents have college and above degrees, and are more likely to have the ability to overcome fertility barriers. In addition, the positive effect of family support on perceived behavioral control may be offset by the negative effect of perceived risk. When p > 0.05, hypothesis H6 is not tested, indicating that there is no significant impact of policy support on perceived behavioral control because the questionnaire focuses on measures of fertility policy awareness and promotion.In terms of perceived behavioral control, the p values of economic support, family support, policy support, and public support in relation to behavioral attitudes are all less than 0.05, and the standardized path coefficients are all greater than 0, indicating that economic support, family support, policy support, and public support have a significant positive effect on behavioral attitudes; therefore, the original hypotheses H8, H9, H10, and H11 are tested. The standardized path coefficient (0.463) is the largest, indicating that the economy has the greatest influence on behavioral attitudes.In terms of behavioral attitudes, the Based on the Theory of Planned Behavior, this paper constructs a structural equation model of the factors influencing women’s fertility intentions, and obtains the theoretical model shown in Overall, economic support and public support have a significant positive effect on perceived behavioral control. Economic support significantly impacts perceived behavioral control and public support negatively impacts perceived behavioral control, while family support and policy support do not have significant impacts on it. Economic support, family support, policy support, and public support have a significant positive effect on behavioral attitudes, in which economic support has the largest effect, followed by economic support, family support, policy support, and public support. Perceived behavioral control and behavioral attitude impact fertility intentions positively, in which behavioral attitude has the larger effect. The results are shown as follows.(1) The role of economic support in perceived behavioral controlThe standardized path coefficient of economic support affecting perceived behavioral control is 0.249, which means that if economic support increases by one standard deviation, perceived behavioral control will increase by 0.249 standard deviations. It can be seen from (2) The role of perceived risk in perceived behavioral controlThe standardized path coefficient of perceived risk affecting perceived behavioral control is −0.251, which means that if perceived risk increases by one unit, perceived behavioral control will be reduced by 0.251 units. Perceived risk during the epidemic contains many elements; for example, Liu et al.  pointed (3) The role of family support in perceived behavioral controlFamily support includes three parts: support from family members, family members’ situation, and family living environment. Perceived behavioral control is the degree to which individuals perceive their fertility behaviors to be controlled. Individuals consider the ability, resources, and opportunities they have to give birth, as well as the health status of the couple, job opportunities, economic situation, and the possibility that a babysitter can help them take care of the child. When there are more relevant resources and opportunities, the perceived behavioral control and fertility intentions are stronger. For example, Lin et al.  confirmeDuring this period, respondents perceived more risks, such as health and unemployment risks. We suggest in (4) The role of public support and policy support in perceived behavioral controlThe standardized path coefficient of public support affecting perceived behavioral control is 0.211, which means that if public support increases by one unit, perceived behavioral control will increase by 0.211 units. Adequate labor rights protection and high-quality public service provision are important to easing the family–career conflicts of working women and improving their beliefs about birth control. Sound job security and social care resource support help boost the fertility intentions of women of childbearing age in China. This conclusion is the same as in the study of Tang and Li , which p(5) The role of economic support, family support, policy support and public support in behavioral attitudesThe standardized path coefficient of economic support affecting behavioral attitudes is 0.463, which means if economic support increases by one unit, behavioral attitudes will increase by 0.463 units. The standardized path coefficients of family support, policy support, and public support affecting behavioral attitudes are 0.227, 0.238, and 0.200, respectively, which are significantly smaller than 0.463, indicating that the effect of economic support on behavioral attitudes is the largest. In other words, the economic recession caused by COVID-19 has reduced their expectations and happiness regarding future births, increased their insecurity, and thus reduced their willingness to have children .(6) The role of perceived behavioral control and behavioral attitudes in fertility intentionsThe standardized path coefficient of perceived behavioral control affecting fertility intentions was 0.232, which means if perceived behavioral control increases by one unit, fertility intentions will increase by 0.232 units. The standardized path coefficient of behavioral attitudes affecting fertility intentions is 0.614, meaning if behavioral attitudes increase by one unit, fertility intentions will increase by 0.614 units, which is significantly greater than 0.232, indicating that behavioral attitudes have the greatest effect on fertility intentions. However, relevant studies have shown that perceived behavioral control has the greatest impact on behavioral intention , which iThis paper investigates the impact of the COVID-19 epidemic on the fertility intentions of females by distributing questionnaires, constructing structural equation models based on the Theory of Planned Behavior, using economic support, perceived risk, policy support, public support, and family support as external variables, selecting perceived behavioral control and behavioral attitude as determinants of fertility intentions, and using fertility intentions as the outcome variable to construct a structural equation model to explore female fertility intentions during the epidemic. Based on theoretical analysis and empirical research, the main findings are summarized and corresponding policy recommendations are proposed, including enhancing women’s fertility well-being and increasing fertility intentions, to achieve a fertility policy to regulate the population, delay aging in China, and optimize the population structure.(1) Epidemic negatively affects women’s fertility intentionsAfter the outbreak of COVID-19, most women were pessimistic about having children, with only 27% of respondents planning to have children in the next three years. In addition, most women who had plans to have children before COVID-19 postponed or canceled their plans to varying degrees, with 31.5% of respondents planning not to have any more children and only 19.15% of respondents not changing their plans.(2) Perceived behavioral control and behavioral attitudes positively impact fertility intentionsDuring the epidemic, perceived behavioral control and behavioral attitudes positively impacted women’s fertility intentions to varying degrees, with behavioral attitudes having the greatest effect. The economy has had the greatest impact on perceived behavioral control and behavioral attitude, and women’s career development and psychological factors have a significantly negative impact on perceived behavioral control, indicating that most women are still unable to solve their economic challenges, and career development and psychological problems are still important factors that hinder the female population.(1) Provide financial support to reduce the cost of childbirth for womenAccording to the results of this paper, economic pressure is still one of the most important factors affecting women’s fertility intentions. As the main body of policy formulation and implementation, the government should implement a series of policies to reduce families’ economic pressures so as to improve female fertility intentions. First, granting maternity benefits for different maternity situations, such as first and second birth, can be encouraged. Secondly, improving maternity insurance policies, raising the level of maternity insurance coordination and insurance levels, and especially raising the reimbursement ratio for childbirth expenses and the level of maternity benefits, may be adopted. Third, regulating housing prices to a reasonable level, implementing preferential housing policies, providing different amounts of housing allowance for families with two and three children based on the family’s economic situation and the different circumstances of childbirth, lowering mortgage interest rates to reduce the direct cost of raising a family, and providing more financial support to women can also be implemented.(2) Eliminate employment discrimination and alleviate women’s fertility anxietyFemale employment discrimination still exists, and the implementation of the “third child” policy will mean women encounter even more severe employment challenges. In accordance with the actual situation in each province and city, the government should further optimize the protection of female employment rights, implement maternity tax incentives to enterprises employing women who give birth, and speed up the construction of a reasonable and effective mechanism for sharing maternity costs among the state, enterprises, and families, while reducing the employment costs of enterprises and preventing them from transferring employment costs to working women, thereby solving women’s employment problems. Female employees should enjoy equal rights to men at work, and the government should improve the Labor Law and the Labor Contract Law. Companies that deprive female employees of rights and interests should be punished, and made to implement the policy thoroughly. The provisions and regulations for protecting the rights and interests of women wishing to have three children should be optimized and updated promptly to further improve the protection of women’s rights and interests.(3) Improve public services and optimize education allocationMost working women are struggling with balancing family and work given their limited time and energy, while the various forms of public services and infrastructure in the community do not meet their demands. Therefore, the government should adjust the construction of investment in public service resources and facilities, optimize the allocation of education, increase the construction of public childcare services, and provide financial support to improve the quality of existing childcare institutions. In addition, the government should build new public institutions and increase the number of public day childcare institutions and nurseries in places without enough childcare institutions. By providing financial subsidies to communities, the government should be able to bring the community into full play, integrate community resources, set up community-wide childcare centers (where children can be cared for by the employees of the centers), and promote diversified childcare services to meet different childcare needs.Our study of the mechanisms affecting fertility intentions is comprehensive, and the intrinsic effects and action mechanisms are complex. This paper only selects some representative factors and indicators for analysis, without discussing in depth whether there is an indirect influence mechanism. It should subsequently look for more influencing factors, refine the content of the study, and carry out systematic research around this area in order to improve the accuracy of the research results . Since t


Currently the five-year survival of childhood cancer is up to 80% due to improved treatment modalities. However, the majority of childhood cancer survivors develop late effects including infertility. Survivors describe infertility as an important and life-altering late effect. Fertility preservation options are becoming available to pre- and postpubertal patients diagnosed with childhood cancer and fertility care is now an important aspect in cancer treatment. The use of fertility preservation options depends on the quality of counseling on this important and delicate issue. The aim of this manuscript is to present a questionnaire to determine the impact of fertility counseling in patients suffering from childhood cancer, to improve fertility care and evaluate what patients and their parents or guardians consider good fertility care.Within the framework of the EU-Horizon 2020 TREL project, a fertility care evaluation questionnaire used in the Netherlands was made applicable for international multi-center use. The questionnaire to be used at least also in Lithuania, incorporates patients’ views on fertility care to further improve the quality of fertility care and counseling. Results evaluate fertility care and will be used to improve current fertility care in a national specialized pediatric oncology center in the Netherlands and a pediatric oncology center in Lithuania.An oncofertility-care-evaluation questionnaire has been developed for pediatric oncology patients and their families specifically. Results of this questionnaire may contribute to enhancement of fertility care in pediatric oncology in wider settings and thus improve quality of life of childhood cancer patients and survivors.The online version contains supplementary material available at 10.1186/s12885-022-09450-2. Currently the five-year survival rate of childhood cancer is up to 80% in most European countries due to improved treatment regimens , 2. HoweAdequate fertility counseling for girls with cancer comprises of individualized future fertility risk assessment and communication as well as provision of strategies to preserve gonadal material in order to maintain maximal fertility potential. This has been integrated in the Dutch amendment of the Edinburgh criteria “Standard of Cancer Care for fertility preservation” –29. New Preserving ovARian function through cryoprEservation and informing girLs with cancer about infertility due to gonadotoxic treatment (PAREL) study and the TREL initiative. The aim of this manuscript is to present an oncofertility-care-evaluation questionnaire. The questionnaire aims to determine the impact of receiving information and fertility counseling in childhood cancer patients and their parents/guardians and evaluate what they consider good fertility care. This insight may be used to improve fertility care.However, it is unknown how patients experience the fertility care and to date no validated questionnaires exist to evaluate this in a pediatric cancer setting. We intent to improve oncofertility care and evaluate what patients consider adequate fertility care including the impact of receiving information regarding reproductive health and fertility counseling towards fertility preservation in childhood cancer patients. Both onco-fertility care and fertility preservation methods for girls are considered standard of care since publication of the ASRM statement and IGHG guidelines , 36, 37.Voortplanting Bezorgdheid schaal)) by Garvelink et al. [The questionnaire for evaluation of fertility care for girls, currently used in the Netherlands, is based on multiple validated questionnaires concerning decision regret, reproduction concern and the evaluation of fertility care in an adult setting. Relevant sections of these questionnaires were combined in the new questionnaire. Additionally, some questions were amended to fit the pediatric oncology setting and some new questions were developed. The questions from the Decision regret scale by Brehaut et al.  and the k et al.  was usedk et al. .The questionnaire is divided in 5 sections. The first section includes general questions to evaluate how worried patients and parents were about fertility at the time of diagnosis, whether they could recall having a conversation about fertility, and whether they proactively asked for this information. The second section contains questions concerning the first conversation regarding fertility with the nurse practitioner or the pediatric oncologist and focusses on timing and clarity of the information. The third section contains questions regarding the counseling with respect to timing and content, the knowledge on the personal risk of gonadal damage, as well as risks and benefits of the fertility preservation options. Questions regarding emotions of the patients and parents and feelings of control are also included. The fourth section contains questions regarding perceived knowledge on infertility following the information and emotions concerning the information. The last section consists of 4 open questions regarding improvement of fertility care.The initial questionnaire was developed at the Princess Máxima Center and contains 41 items. It is given to all girls who received counseling by a fertility-gynecologist and participate in the PAREL study. The PAREL study has been approved by the Medical Ethics Committee Utrecht (METC nr. NL72115.041.19). To make it applicable for multicenter use within the TREL framework, and in particular in Lithuania, the questions were translated from Dutch to English and afterwards from English to Lithuanian , intermediate (4000–6000 mg/m2) or high risk (≥6000 mg/m2) of gonadal damage [Since May 2018 all pediatric cancer care has been centralized in one national pediatric cancer center, the Princess Máxima Center. Around 600 children are newly diagnosed with pediatric cancer in the Netherlands every year. A 5-step oncofertility care plan is implemented since 2019 . These 5l damage . HoweverThe TREL consortium is formed by VULSK and 8 leading research institutions each covering different areas of the project activities according to their expertise in pediatric oncology. TREL will be delivered in 7 work packages (WP) addressing training in tumour specific laboratory research and clinical trials, cross-cutting education on genome-wide sequencing and treatment innovations, enhancing skills in observational studies on the quality of survivorship including fertility preservation and research methodology as well as project and innovation management. TREL is a European twinning effort that aims to strengthen research networking and education in Lithuania with the ultimate goal to improve survival and quality of life of children with solid tumours . The development of the questionnaire is part of WP6 of the TREL collaboration. WP6 specifically focusses on the quality of survivorship and late effects research.In Lithuania the questionnaire will be implemented at the Center for Pediatric Oncology and Hematology (CPOH) at VULSK, which is the biggest pediatric oncology and hematology center in Lithuania and the Baltic region. VULSK covers two thirds of pediatric cancer patients in Lithuania. Children aged from 1 month to 18 years are treated at the VULSK, every year 50–60 new patients with childhood cancer are diagnosed and treated. Approximately 50 patients and 20 survivors are currently in treatment or in remission for less than 5 years and are regularly followed up at the VULSK. At the moment, fertility counseling at VULSK is rather sporadic, gonadal tissue preservation is available after a consultation with qualified fertility specialists, but there is no developed fertility care system in place. In Lithuania the preservation of reproductive tissue is embedded in the national legislation and can be offered only to children over 14 years old. A triage system similar to the one used in the Princess Máxima Center is being developed to stratify patients according to their risk for infertility/gonadal damage . PatientThe increasing number of CCSs is a reason why research is increasingly focusing on their well-being. They are at risk for infertility, which affects quality of life. As reported in a previous study on reproductive health of Lithuanian CCSs , many ofIt is well known that patients and parents do not remember all of the given information in stressful situations. Some studies even suggest that only 20% of the given verbal information is retained , 44. In Since fertility care is structured differently in the Netherlands and Lithuania, the Lithuanian questionnaire was adjusted to the local situation, e.g. a nurse practitioner is not available in the Lithuanian health system. Also the patient population will be slightly different, since a proportion of VULSK patients who receive a questionnaire may not have received oncofertility counseling by experts. In comparison, all patients receiving the questionnaire in the Netherlands have received fertility counseling from fertility experts. Bearing in mind the different cultural backgrounds, different legislations and different system of fertility counseling of childhood cancer patients in two different countries, it could be expected that the answers to the same questions may vary. This may reveal cultural differences that may influence future fertility care strategies.Oncofertility counseling is an important part of pediatric cancer care, yet no questionnaire to evaluate this existed for the pediatric population. The developed questionnaire to evaluate oncofertility care in two countries may provide insight in the views of patients and their family on offered fertility care and on improvements that could be made. Results of this questionnaire may contribute to enhanced oncofertility settings in pediatric oncology departments in a wider range of cultural and geographic settings, thereby improving quality of life of childhood cancer patients and survivors.Additional file 1.


In response to CXCL12, CXCR4 and ACKR3 both recruit β-arrestin 2, regulating the assembly of interacting proteins that drive signaling and contribute to the functions of both receptors in cancer and multiple other diseases. A prior proteomics study revealed that β-arrestin 2 scaffolds pyruvate kinase M2 (PKM2), an enzyme implicated in shifting cells to glycolytic metabolism and poor prognosis in cancer. We hypothesized that CXCL12 signaling regulates PKM2 protein interactions, oligomerization, and glucose metabolism. We used luciferase complementation in cell-based assays and a tumor xenograft model of breast cancer in NSG mice to quantify how CXCR4 and ACKR3 change protein interactions in the β-arrestin-ERK-PKM2 pathway. We also used mass spectrometry to analyze the effects of CXCL12 on glucose metabolism. CXCL12 signaling through CXCR4 and ACKR3 stimulated protein interactions among β-arrestin 2, PKM2, ERK2, and each receptor, leading to the dissociation of PKM2 from β-arrestin 2. The activation of both receptors reduced the oligomerization of PKM2, reflecting a shift from tetramers to dimers or monomers with low enzymatic activity. Mass spectrometry with isotopically labeled glucose showed that CXCL12 signaling increased intermediate metabolites in glycolysis and the pentose phosphate pathway, with ACKR3 mediating greater effects. These data establish how CXCL12 signaling regulates PKM2 and reprograms cellular metabolism. Chemokine receptor CXCR4 and the atypical chemokine receptor ACKR3 (formerly CXCR7) regulate essential processes in normal physiology and numerous diseases, including cancer, atherosclerosis, neurodegeneration, and autoimmunity . While bA prior mass-spectrometry-based analysis found more than 100 different proteins that interacted with both β-arrestins. Additional proteins selectively interacted with β-arrestin 1 or 2 . PrelimiBoth CXCR4 and ACKR3 signaling pathways promote tumor growth and metastasis in breast cancer and multiple other malignancies, making these receptors potential targets for therapy and molecular imaging ,13,14,15Using cell-based and in vivo imaging combined with mass spectrometry analyses, we discovered that CXCL12 signaling through CXCR4 and ACKR3 regulates interactions between ERK2, β-arrestin 2, and PKM2. Both receptors reduce PKM2’s association with β-arrestin 2 and decrease the oligomerization of this enzyme, a characteristic shift in aerobic glycolysis. We also demonstrated that CXCR4 and, to a greater extent, ACKR3, promote the glucose metabolism through glycolysis and the pentose phosphate pathway, processes associated with the proliferation of cancer cells. These results advance the understanding of how CXCL12, CXCR4, and ACKR3 regulate intracellular signaling and highlight the functions of these pathways in shaping metabolism.2. We renewed cultures from frozen stocks at least every three months. We generated spheroids of MDA-MB-231 cells and human mammary fibroblasts as described previously . Th. Thp < 0 < 0.05) F. We als < 0.05) G, consisWe compared how CXCL12 signaling through CXCR4 versus ACKR3 regulates PKM2 oligomers. We stably expressed complementation reporters PKM2-CBRN/PKM2-CBC in MDA-MB-231 cells transduced with CXCR4 or ACKR3 A. We tre13C6 glucose for 15 min before measuring labeled metabolites using mass spectrometry. As compared with vehicle control, cells treated with CXCL12 showed lower amounts of glucose-6-phosphate/fructose-6-phosphate and increased intermediates in glycolysis: fructose-1,6-bisphosphate (FBP) and 2-phosphoglycerate/3-phosphoglycerate  to counter oxidative stress . In cellhosphate A,B. The 13C6 glucose revealed that CXCL12 signaling through CXCR4 and ACKR3 promotes the glucose metabolism through the pentose phosphate pathway and the accumulation of intermediate metabolites in glycolysis. While both CXCR4 and ACKR3 drive these protein interactions and metabolic shifts, we noted differences in the kinetics and magnitude of signaling responses and metabolites. CXCR4 triggered the greater induction and more rapid recruitment of β-arrestin 2 to the receptor, and the association between ERK2 and β-arrestin 2. By comparison, ACKR3 sustained interactions with β-arrestin 2 even after CXCL12 removal and produced greater changes in PKM2 oligomerization state and glucose metabolism. The causes of these differences in kinetics and outputs have yet to be fully defined. Nevertheless, this study establishes a previously unknown function of CXCL12 signaling to regulate PKM2 and glucose metabolism, connecting CXCR4 and ACKR3 to the metabolic adaptations of cancer cells.Building on our past work with luciferase complementation systems for imaging protein interactions in living cells, we established a unique suite of reporters to investigate intracellular signaling complexes regulated by CXCR4 and ACKR3. Cells with a high demand for the synthesis of nucleic acids, particularly cancer cells, preferentially express PKM2 and maintain the enzyme as a dimer . The lowThe scaffolding function of β-arrestin 2 potentially regulates other reported mechanisms of action for PKM2 in metabolism, as well as tumor progression. CXCL12 stimulates the recruitment of ERK2 to β-arrestin 2 and the formation of multi-protein interactions, including PKM2, which may facilitate the phosphorylation of PKM2, as detected downstream of CXCL12-ACKR3 signaling. The ERK phosphorylation of PKM2 is essential for aerobic glycolysis in cancer cells in cell-based assays and mouse models of cancer . A priorThe available evidence supports a strong correlation between the increased expression of PKM2 in cancer and the metabolic adaptations necessary for proliferation. However, ongoing studies also suggest complex, possibly context-dependent effects of PKM2 in cancer. Nuclear PKM2 has been reported as a protein kinase, phosphorylating histones and other epigenetic regulators to increase the expression of oncogenic proteins such as Myc- and cyclin-dependent kinases . The proOverall, the current study reveals that CXCL12 signaling through CXCR4 or ACKR3 regulates PKM2 scaffolding on β-arrestin 2 and reduces the oligomerization of this glycolytic enzyme. Since dimeric PKM2 is known to drive the glycolytic metabolism of glucose and accumulation of metabolites in glycolysis and the pentose phosphate pathway, our signaling data provide one potential mechanism for CXCL12-stimulated shifts in the metabolism of breast cancer cells. We studied the signaling effects through only CXCR4 or ACKR3. However, we and others have shown that these receptors form hetero-oligomers with distinct signaling responses . Future


The purpose is to improve the teaching and learning efficiency of college Innovation and Entrepreneurship Education (IEE). Firstly, from the perspective of aesthetic education, this work designs the teacher and student sides of the Computer-aided Instruction (CAI) system. Secondly, the CAI model is implemented based on the weight sharing and local perception of the Convolutional Neural Network (CNN). Finally, the performance of the CNN-based CAI model is tested. Meanwhile, it analyses students’ IEE experience under the proposed CAI model through a case study of Music Majors from Xi’an Conservatory of Music. The experimental data show that the CNN-based CAI model can respond quickly and stably when users access different functional modules, such as webpage browsing. The proposed CAI model increases students’ entrepreneurial interest, skills, and knowledge by 55.62, 57.32, and 72.12%, respectively. Students’ entrepreneurial practice ability has been improved by over 50.00%; such an increase in entrepreneurial practice ability has also shown individual differences. Thus, the proposed Music Majors-oriented IEE-infiltrated CAI model based on CNN improves students’ entrepreneurial practice ability and reflects the positive experience of Music Majors on IEE. The finding provides references for the step-by-step identification of the CNN-based CAI model and has certain guiding significance for analyzing the effect of college IEE. As China’s social progress accelerates, cultivating innovative, application-oriented, and entrepreneurial talents has been put on the domestic higher educational agenda . HoweverOn the other hand, music talents are special groups with exceptionally well-talented music skills and performance techniques. Therefore, infiltrating IEE into music education helps Music Majors fully display their talents. It gives students more freedom to think, judge, empathize, imagine, and express their feelings through music . Thus, ICurrently, IEE in Chinese CAUs lacks an effective practice platform, and the instruction system is devoid of utility. This work aims to improve the effectiveness of college IEE and fill the research gap of a versatile and effective instruction system in the IEE development. Firstly, from the perspective of aesthetic education, the teacher side and student side of the CAI system are designed. Secondly, the CAI model is implemented based on CNN’s weight distribution and local perception. The innovation is to test the performance of the proposed CAI system on the CNN framework by investigating and analyzing the IEE experience of Music Majors from Xi’an Conservatory of Music. The logical structure is shown as follows. In Chapter 1, the background of CNN and CAI is introduced. Chapter 2 introduces the concepts of the CAI system and IEE. Then, in Chapter 3, the performance of the proposed CAI system is demonstrated. Chapter 4 analyses the data results and draws the research conclusions. This work can provide some references for designing and optimizing CAI systems through advanced technologies like DL. It also contributes to integrating IEE with other subjects to improve students’ Entrepreneurial Spirit (ES) and learning effect.The traditional college music education in China fails to consider the multicultural environment in curriculum design. It cannot arouse students’ music learning or composition initiative. Students learning interest in the music is lost. Besides, Music Majors are special students and rely more on practice than theoretical learning, which has not been fully reflected in the existing music education modes . Most ofAs shown in In According to m*n convolution kernel on an original image X reads:Neural network models simulate the structure and behavior of the biological neuron system for distributed and parallel information processing. It can process information by adjusting the weight between neurons. In particular, CNN, a feedforward Neural Network (NN) model with several convolution layers and pooling layers, can well lend to image processing . Meanwhi1w, 2w, and wmn are the weights of convolution kernel W. 1x, 2x and xmn are the corresponding pixels on the original image. z is the value of the original image X after convolution. The convolution kernel convolutes and traverses the image using a certain-step sliding window. The sliding window will skip some pixels. Here, the convolution kernel covers 9 pixels in the original image each time. The output of an F-step convolution on an n-size original image reads:O indicates the size of the output image, and s denotes the convolution step. Convolution operations shrink and deform the image. The image edge is formed through one-pixel output, while the image center is formed through multiple overlapping convolution kernels. Thus, some edge pixels will be lost in convolution. The edge pixels in the center of the convolution kernel are extended to the false pixels outside the edge in the convolution kernel. Then, Eq. 3 calculates the size of the output image:p denotes the filled pixel size, and then the image size of the original image isn+2p. Images’ spatial connection is also a local pixel connection: the closer the pixels, the stronger the correlation is. The correlation of distant pixels is weak. Therefore, each neuron does not need to perceive the global image. Global information can be obtained by perceiving local information and integrating it at a higher level. The structure of the visual system inspires the idea of a partial network connection in biology. Neurons in the visual cortex receive information locally . Here, the CNN intelligent sensor identifies the CAI activities under different modes. Learning activity-oriented motion and speech recognition is to classify the accelerometer data sequence recorded by the special strap or smart computer and mobile phone into known and clearly defined motions. College IEE improves students’ quality through the relevant curriculum system, such as the spirit of independence, innovation, adventure, as well as ES. Meanwhile, IEE reshapes students’ employment concepts and entrepreneurial beliefs . The IEEAnalyzing the characteristics of college Music Majors-oriented IEE reveals that Music Majors’ cognition and entrepreneurship experience are subjective. The teaching process is exploratory. The efficiency of teaching activities needs to be improved, and the practice link needs to be re-examined.Through the step-by-step implementation of the CAI system from the perspective of CNN and aesthetic education, 108 Music Majors in a college are randomly selected for a questionnaire survey (QS). Then, the invalid QSs, such as missed selection and multiple selections, are picked out and eliminated, with 98 valid ones reaching a 90.7% recovery rate. In order to verify the data reliability, Statistical Packages for Social Science (SPSS) and Cronbach’s α coefficient are used to analyze the reliability and validity of the QS. The data results show that the correlation coefficients of each dimension are higher than 0.7, indicating a good structural validity. The reliability of the QS is also analyzed. Lastly, the formal QS in the study is prepared according to the standard. The measured Cronbach’s α is between 0.75 and 0.90, which significantly conforms to the reliability range. The structure of the QS is given in Here is the specific experimental environment:64-bit Windows10 operating system; an Intel (R) Core (TM) i7-8700K 3.7 GHz Central Processing Unit (CPU); a 16GB Random Access Memory (RAM); and the NVIDIA GeForce GTX 1080Ti 8GB Graphics Processing Unit (GPU). Through the comparative analysis before and after using the proposed CNN-based CAI system, this work analyses the practical ability of Music Majors’ IEE. The CLPsyh2017 ReachOut dataset is also selected to train and test the CNN. The dataset includes 1,188 pieces of test data and 400 pieces of training data. The test and training data are sorted out to train and test the model, respectively.n = 500. First, the open CAI system interface is closed, and users log in synchronously. When concurrent user number = 57, the performance of the proposed CNN-based CAI system is analyzed and tested. This section verifies the proposed CNN-based CAI model’s different module performance. The model training iteration Subsequently, a QS is conducted on 108 Xi’an Conservatory of Music students. The proposed CNN-based CAI model is introduced in college IEE. The growth coefficient is used to study the changes in Music Majors’ entrepreneurial interest, entrepreneurial skills, and entrepreneurial knowledge within 1 month. As in In social progress and economic development, innovative and entrepreneurial talents have always been the main driving force. This work proposes a college Music Majors-oriented IEE-infiltrated CAI model based on CNN from the perspective of aesthetic education. It chooses Music Majors from Xi’an Conservatory of Music using the proposed CAI model to study their increase in practical abilities. The results show that with 57 concurrent users, the average response time of the proposed CAI model is between 0.19 and 0.28 s. The proposed CAI model can respond to 2.67–5.65 clicks/second for user web browsing, which can meet the user’s practical needs of 2.38–5.67 clicks/second. Thus, the proposed CAI model has a quick and stable response time for different modules. Music Majors’ entrepreneurial interest, skills, and knowledge have increased by 55.62, 57.32, and 72.12%, respectively, after introducing the proposed CAI model. Music Majors’ average practical entrepreneurial ability before and after introducing the proposed CAI model is 0.28 and 0.60, respectively. Therefore, the proposed Music Majors-oriented IEE-infiltrated CAI model based on CNN can improve Music Majors’ learning experience for IEE, entrepreneurial interest, skills, knowledge, and practice abilities. The proposed CAI model can provide some experimental reference for researching the influencing factors of college IEE. Last but not least, there are still some deficiencies. The QS is issued from the perspective of Music Majors, and the experience analysis of CAI and IEE is not comprehensive. In future research, it is prospected to expand the major and numbers of student samples and analyze the experience of college IEE from multiple perspectives. The aim is to provide some theoretical support for the development of CNN and college IEE.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by the Hunan City University Ethics Committee. The patients/participants provided their written informed consent to participate in this study. Written informed consent was obtained from the individual(s) for the publication of any potentially identifiable images or data included in this article.The author confirms being the sole contributor of this work and has approved it for publication.The author declares 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.


The COVID-19 pandemic has affected various dimensions of the economies and societies. At this juncture, this paper examines the effects of pandemics-related uncertainty on fertility in the panel dataset of 126 countries from 1996 to 2019. For this purpose, the World Pandemics Uncertainty Indices are used to measure the pandemics-related uncertainty. The novel empirical evidence is that pandemics-related uncertainty decreases fertility rates. These results are robust to estimate different models and include various controls. We also try to explain why the rise in uncertainty during the COVID-19 pandemic has resulted in the fertility decline. The COVID-19 pandemic has affected various aspects of the economy and society. It has led to different fiscal and monetary policy implications , which aFertility is also affected by cultural, economic, political, and social factors . PreviouIt is also important to note that uncertainty shocks can also be essential in fertility behavior. Expectations related to business cycles can affect the change in fertility rates. For instance, in a recent empirical study on more than 100 million births in the United States from 1988 to 2014, Buckles et al.  observedAll in all, families can postpone having a child during periods of higher uncertainty . We suggIn this paper, we assume that pandemic-related uncertainty by a decline in consumer confidence and reduced or delayed household consumption–suggesting that “precautionary saving motivation” is the primary mechanism affecting fertility plans during the pandemic. However, we can also indicate that becoming pregnant does not have to immediately increase or affect consumption; instead, having a child influences family consumption and spending over a very long period when the child is growing up and in education. In addition, our precautionary saving perspective is reducing the potentially wide-ranging impact of uncertainty just to one specific factor, ignoring many other relevant motivations and potential drivers of fertility decisions, including the actual experiences of economic hardship , health consequences of infection or limitations due to lockdowns and government measures intended to limit the spread of the COVID-19. For instance, women in some Latin American countries were advised not to get pregnant during the Zika Epidemic as the disease could damage the fetus's brain during pregnancy. Thus, precautionary saving should be seen as one of many possible factors affecting fertility due to pandemic-related uncertainty.This paper aims to contribute to the empirical literature by examining the effects of pandemics-related uncertainty on fertility behavior. We focus on precautionary savings motivation to explain the level of fertility rate and changes. At this point, we use a novel measure of pandemics-related uncertainty, so-called the World Pandemics Uncertainty Index (WPUI), proposed by Ahir et al. . The WPUThe rest of the paper is organized as follows. Section Literature review on fertility decisions reviews the previous articles in the literature. Section Empirical model and data sets the empirical model and provides the features of the data. Section Empirical results discusses the empirical findings for the countries at different stages of economic development. Section Robustness checks provides the results of the robustness analyses based on other empirical models. Section Conclusion and recommendations concludes.The fertility theory documents a negative association between fertility and economic development in the long run. Specifically, fertility rates decline as a country develops , 13, 25.In microeconomics, the marginal utility of consumption is convex , 28. ThiMany studies have provided evidence of the negative impact of human capital on fertility rates in the long run –32. EarlThere is also a negative theoretical relationship between the fertility rate and per capita income. According to Greenwood and Seshadri , the negIn addition, Ehrlich and Kim , AlhassaFollowing these discussed papers, we should include economic development, human capital, and life expectancy as the main controls in the empirical models for the long run.There are also several determinants of the changes in the fertility rates, which are mainly related to short-run business fluctuations. Uncertainty is one of the leading determinants of fertility changes, given that consumption and income uncertainty decrease fertility rates. According to the precautionary motivation approach, uncertainty in income leads to pessimist expectations. It decreases “consumer confidence” during recessions . ConsumePrevious empirical papers suggest finding a “purely exogenous” measure of uncertainty to investigate the relationship between uncertainty and fertility. We offer that the WPUI is a purely exogenous indicator to measure uncertainty.Several papers have focused on the effects of the COVID-19 pandemic on fertility rates. However, previous articles focus on the impact of the earlier pandemics before COVID-19. For instance, Boberg-Fazlic et al.  documentRegarding the COVID-19 studies, Aassve et al.  discuss On the other hand, Ullah et al.  discuss Furthermore, Rovetta  demonstrvia declining trade volumes and portfolio flows. Pandemics also have created uncertainty over future income, affecting the expectations similar to business cycles. We aim to contribute to the empirical literature by connecting pandemics-related uncertainty and the fertility rate. Following De la Croix and Pommeret  to (4), ΔThe dependent variables are the level and the change in total fertility rate (births per woman), downloaded from World Bank . Some moThe primary variable of interest is the level and the change of the WPUI in the lagged and the current forms. The related data have been accessed on the Ahir et al.  introducColumns 1 and 2 report the results for all (126) countries from 1996 to 2019. The first column provides the findings of the current WPUI, and the second column provides the results of the lagged WPUI. In addition, the findings of 92 non-OECD economies are provided in Columns 3 and 4. The results of 34 OECD economies are reported in Columns 5 and 6.In all countries, the coefficient of the current WPUI is −0.195, and it is significant at the 1% level. Similarly, the coefficient of the current WPUI is −0.166 for the non-OECD economies, and it is significant at the 1% level. However, the current WPUI is −0.566 for the OECD economies, statistically insignificant. Note that the coefficients of the lagged WPUI are positive, but they are statistically insignificant. These results indicate that the pandemics-related uncertainty has an adverse and temporary effect on fertility behavior in developing countries. This evidence may be explained that the non-OECD economies have less human capital than the OECD economies. The evidence can also be related to the issue of economic resources and welfare state policy implications in the OECD economies, and these countries are not significantly affected by uncertainty shocks related to pandemics.Furthermore, the control variables' significant effects on the fertility rates. For instance, the lagged per capita GDP is positively associated with the fertility rate. The related coefficients are statistically significant at the 5% level at least. The lagged life expectancy decreases the fertility rate, and their coefficients are also statistically significant at the 1% level. The lagged human capital is positively related to the fertility rate. The coefficients are statistically significant at the 1% level for all countries and non-OECD economies; however, the coefficients are statistically insignificant in OECD economies. This evidence suggests that the effects of pandemics shocks on fertility are essential even though various controls are included.Our findings are based on the coefficients of the level of WPUI, and the lagged WPUI shows no significant effect on fertility. This evidence related to the issue that the impact of the uncertainty shock on fertility behavior is mainly valid in the short run, as previous papers e.g., , 66] sug sug66)] Again, Columns 1 and 2 provide the findings for all (126) countries from 1996 to 2019. The first column reports the results of the current WPUI, and the second column provides the findings of the lagged WPUI. In addition, the results of 92 non-OECD economies are reported in Columns 3 and 4. The results of 34 OECD economies are compared in Columns 5 and 6.In all countries, the coefficient of the current WPUI is −0.285, and it is significant at the 1% level. Similarly, the coefficient of the current WPUI is −0.243 for the non-OECD economies, and it is significant at the 1% level. Also, the current WPUI is −0.82 for the OECD economies and is statistically significant at the 1% level. In addition, the coefficients of the lagged WPU are negative, but they are statistically insignificant. These findings show that pandemics-related uncertainty has an adverse and temporary effect on the fertility rate in all countries.On the other hand, the significant effects of the controls on the fertility rates are observed. Specifically, the lagged per capita GDP is positively related to the fertility rate. Their coefficients are statistically significant at the 1% level. The lagged life expectancy reduces the fertility rate, and the related coefficients are also statistically significant at the 1% level. The lagged human capital increases the fertility rate. The corresponding coefficients are statistically significant at the 1% level for all countries and the non-OECD economies; however, the coefficients are statistically insignificant in the OECD economies. These results show that pandemics hurt fertility, mostly in poor or developing economies. These findings indicate that the effects of pandemics shocks on fertility are crucial when various controls are included. According to most papers in the empirical literature, the negative impact of COVID-19 is mainly valid in developed economies. However, according to Aassve et al. , the COVAgain, Columns 1 and 2 report the results for 126 countries from 1996 to 2019. The first column provides the findings of the current WPUI. The second column reports the results of the lagged WPUI. In addition, the results of 92 non-OECD economies are provided in Columns 3 and 4. In comparison, the findings of 34 OECD economies are reported in Columns 5 and 6.In all countries, the coefficient of the current WPUI is −0.356, and it is significant at the 1% level. Similarly, the coefficient of the current WPUI is −0.316 for the non-OECD economies, and it is significant at the 1% level. However, the current WPUI is −0.787 for the OECD economies, which is statistically insignificant. Moreover, the coefficients of the lagged WPU are positive, but they are statistically insignificant. These results indicate that the pandemics-related uncertainty has an adverse and temporary effect on fertility behavior in developing economies.It is also noteworthy that the controls statistically affect the fertility rate. Specifically, the lagged fertility rate is statistically significant at 1%, indicating a substantial persistency in fertility decisions. The lagged per capita GDP is positively related to the fertility rate. The related coefficients are statistically significant at the 1% level. The lagged life expectancy decreases the fertility rate, and the associated coefficients are also statistically significant at the 1% level. The lagged human capital spurs the fertility rate. The related coefficients are statistically significant at the 1% level for all countries and the non-OECD economies; however, the coefficients are statistically insignificant in the OECD economies. These results show that the effects of pandemics shocks on fertility are essential when various controls are included.Again, Columns 1 and 2 provide the findings for 126 countries from 1996 to 2019. The first column provides the results of the current WPUI. The second column reports the findings of the lagged WPUI. In addition, the findings of 92 non-OECD economies are reported in Columns 3 and 4. The results of 34 OECD economies are compared in Columns 5 and 6.In all countries, the coefficient of the current WPUI is −0.26. The current WPUI is −0.223 for non-OECD and −0.765 for OECD economies. All of these coefficients are statistically significant at the 1% level. Furthermore, the coefficients of the lagged WPU are positive, but they are statistically insignificant. These findings show that the pandemics-related uncertainty has an adverse and temporary effect on fertility decisions in all countries.It is also noteworthy to note that the significant effects of the controls on the fertility rates are found. For instance, the lagged fertility rate is statistically significant at 1%, meaning a considerable persistency in fertility behavior. The lagged per capita GDP is positively related to the fertility rate. The related coefficients are statistically significant at the 5% level at least. The lagged life expectancy reduces the fertility rate, and the associated coefficients are also statistically significant at the 1% level. The lagged human capital increases the fertility rate. The related coefficients are statistically significant at the 1% level for all countries and the non-OECD economies; however, the coefficients are statistically insignificant in the OECD economies. These findings again indicate that the effects of pandemics shocks on fertility are essential even though various controls are included.In short, various empirical model estimations indicate that pandemics-related uncertainty decreases the fertility rate, and the effect is temporary. Per capita GDP and human capital increase fertility, but life expectancy is negatively associated with fertility. These results are statistically significant for developing economies in all models.The results in This paper examines the effects of pandemics-related uncertainty on fertility behavior in the panel dataset of 126 countries from 1996 to 2019. For this purpose, we use the WPUI introduced by Ahir et al.  to captuOur evidence from the WPUI may also explain why the rise in uncertainty during COVID-19 resulted in fertility decline. It suggests that fertility is negatively associated with business cycles due to increased uncertainty. However, our results are limited to the panel data across various countries. Note that the sample is the main limitation of our findings since most WPUI values are zero in our case. The COVID-19 pandemic can change the dynamics of uncertainty shocks on fertility at this stage. For instance, Wilde et al.  use GoogIt is also essential to indicate that our findings are valid, with a limited number of control variables included. Therefore, future papers include more control variables in the model estimations. Finally, future articles can use individual microdata to analyse pandemics-related uncertainty shocks' effects on the fertility rate.https://databank.worldbank.org/source/world-development-indicators; https://www.rug.nl/ggdc/productivity/pwt/?lang=en; https://worlduncertaintyindex.com/data/.Publicly available datasets were analyzed in this study. This data can be found here: YW: writing introduction and literature review. GG: collection of the data and empirical estimations. CL: writing the results and conclusion and proofreading. 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.