The present study compared the levels of explicit and intrinsic intention to receive COVID-19 vaccination among sexual minority and heterosexual individuals and examined the association of explicit and intrinsic intentions with sexual orientation. We enrolled 171 sexual minority and 876 heterosexual individuals through a Facebook advertisement. The participants’ explicit and intrinsic intentions to receive COVID-19 vaccination and their risk perception of COVID-19 were measured. We discovered that sexual minority individuals had higher levels of explicit and intrinsic intention to receive vaccination relative to heterosexual individuals. Intrinsic intention was positively associated with explicit intention after the effects of demographic characteristics and risk perception of COVID-19 were controlled for. Sexual orientation did not moderate the association between explicit and intrinsic intentions. The present study determined the relationship between sexual orientation and intention to receive COVID-19 vaccination. The coronavirus disease 2019 (COVID-19) pandemic has greatly affected people’s lives worldwide ,5,6,7,8.Equitable access to safe and effective vaccines is critical to ending the COVID-19 pandemic . At presAlthough these vaccines have been demonstrated to be safe and effective ,34,35,36A common method for assessing people’s intention to receive vaccination is the use of a single-item Likert scale or visual analog scale (VAS) ,44. The Vallée-Tourangeau et al. applied the cognitive model of empowerment (CME) as an alternative method to examine people’s intrinsic intention to receive vaccination against respiratory infectious diseases (RIDs) . The CMEDetermining the factors related to explicit intention to get vaccinated against COVID-19 is essential to the development of intervention programs. In prevention motivation theory ,51, the The present study examined three research questions. First, are the levels of explicit and intrinsic intention to receive COVID-19 vaccination different between sexual minority and heterosexual individuals? Second, is intrinsic intention significantly associated with explicit intention when risk perception is accounted for? Third, are the associations between explicit and intrinsic intention to receive COVID-19 vaccination different between sexual minority and heterosexual individuals? Accordingly, this study hypothesized the following:Hypothesis 1 1)..(HRisk perception and intrinsic intention to receive COVID-19 vaccination is significantly associated with explicit intention.Hypothesis 3 3). to measure the participants’ self-reported intrinsic intention to receive COVID-19 vaccination . The DrVA five-item questionnaire was used to measure risk perception regarding contracting COVID-19 . The fivData on participants’ sexual orientation , gender , age, and education level were collected. The participants were then sorted into sexual minority and heterosexual groups.2 and t tests. The two groups’ explicit and intrinsic intention to receive COVID-19 vaccination and risk perception of COVID-19 were compared using a multivariate analysis of covariance (MANCOVA) in which gender, age, and education level were controlled for.Data analysis was performed using SPSS 24.0 . The sexual minority and heterosexual participants were compared with respect to gender, age, and education level using χp value < 0.05 indicated statistical significance.The association between explicit and intrinsic intention to receive COVID-19 vaccination among sexual minority and heterosexual individuals was examined using multiple regression in which risk perception was controlled for. To examine the moderating effect of sexual orientation, we analyzed the interaction between sexual orientation and intrinsic intention to receive COVID-19 vaccination using multiple regression to determine the association of this interaction term with explicit intention to receive COVID-19 vaccination. A two-tailed 1. However, no difference in risk perception was observed between the two groups.The data on demographic characteristics, explicit and intrinsic intention to receive COVID-19 vaccination, and risk perception are presented in 2 increased from 0.039 in Model I to 0.520 in Model II, indicating that intrinsic intention had a significantly stronger association with explicit intention compared with the variables in Model I. These results supported H2.3.We further analyzed the interactions between sexual orientation, risk perception, and intrinsic intention in a multiple regression to examine the association of this interaction term with explicit intention (Model III). The results revealed that the interactions between sexual orientation and risk perception and between sexual orientation and intrinsic intention were not significantly associated with explicit intention, indicating that sexual orientation did not moderate the association of risk perception and intrinsic intentions with explicit intention to receive COVID-19 vaccination. These results did not support HThe present study found that sexual minority individuals had higher levels of explicit and intrinsic intention to receive COVID-19 vaccination relative to heterosexual individuals. Intrinsic intention was positively associated with explicit intention after we controlled for the effects of demographic characteristics and risk perception of COVID-19. Sexual orientation did not moderate the association between explicit and intrinsic intention to receive COVID-19 vaccination.No differences in the level of risk perception of COVID-19 were observed between sexual minority and heterosexual individuals; however, sexual minority individuals had higher levels of explicit and intrinsic intention to receive COVID-19 vaccination relative to heterosexual individuals. Research reported that compared with heterosexual individuals, sexual minority individuals have greater medical needs and thus interact more with medical care providers ; sexual The present study revealed that intrinsic intention to receive COVID-19 vaccination and risk perception of COVID-19 were positively associated with explicit intention after the effects of demographic characteristics were controlled for and that the associations did not significantly differ between sexual minority and heterosexual individuals. Intrinsic intention accounted for almost half of the variance of explicit intention, indicating that the components of intrinsic intention  influenced individuals’ explicit intention to receive COVID-19 vaccination. These components of intrinsic intention ought to be targeted by intervention programs to improve the public vaccination rate.Risk perception is the personal beliefs about the likelihood of suffering a disease . IndividHowever, intrinsic intention, risk perception, and demographic characteristics accounted for only 52% of the variance for explicit intention, indicating the presence of other factors that might have contributed to participants’ explicit intention to receive COVID-19 vaccination but were not examined in the present study; these other factors should be further investigated.The present study is the first to compare sexual minority and heterosexual individuals’ levels of explicit and intrinsic intention to receive COVID-19 vaccination. The results aid the development of group-specific programs for enhancing individuals’ intention to receive COVID-19 vaccination. This study has several limitations. First, the participants were enrolled through a Facebook advertisement. Although enrolling participants using Facebook allows the researchers to quickly and easily reach out to a high number of prospective participants  during tThe present study found that sexual minority individuals had higher levels of explicit and intrinsic intentions to receive COVID-19 vaccination compared with heterosexual individuals. The development of sexual orientation-specific programs is beneficial for enhancing individuals’ intention to receive COVID-19 vaccination. Intrinsic intention was revealed to be positively associated with explicit intention among both sexual minority and heterosexual individuals. The components of intrinsic intention ought to be targeted by intervention programs to increase the public’s intention to receive COVID-19 vaccination.


Bladder cancer is one of most common malignant urinary tract tumor types with high incidence worldwide. In general, transurethral resection of non-muscle-invasive bladder cancer followed by intravesical instillation of chemotherapy is the standard treatment approach to minimize recurrence and delay progression of bladder cancer. However, conventional intravesical chemotherapy lacks selectivity for tumor tissues and the concentration of drug is reduced with the excretion of urine, leading to frequent administration and heavy local irritation symptoms. While nanomedicines can overcome all the above shortcomings and adhere to the surface of bladder tumors for a long time, and continuously and efficiently release drugs to bladder cancers. The rapid advances in targeted therapy have led to significant improvements in drug efficacy and precision of targeted drug delivery to eradicate tumor cells, with reduced side-effects. This review summarizes the different available nano-systems of targeted drug delivery to bladder cancer tissues. The challenges and prospects of targeted therapy for bladder cancer are additionally discussed. In Western countries, the incidence of bladder cancer is different between genders, and the life time risk of bladder cancer is approximately 1.1 and 0.27% in men and women, respectively . BC is rin situ) , the latin situ) . Transurin situ) . Owing tThus, reducing the recurrence rate and preventing tumor progression remain significant challenges in the field of BC research. The gold standard treatment for superficial BC is complete transurethral resection and postoperative adjuvant intravesical instillation of chemotherapy to reduce recurrence and delay or even prevent progression to muscle-invasive disease. Clinical experience supports the utility of intravesical instillation therapy as an auxiliary method to prevent postoperative tumor recurrence. The drug contacts the bladder mucosa via this system to directly or indirectly kill the remaining tumor cells in the bladder, prevent tumor cell planting and reduce the recurrence rate.Bacillus Calmette-Guérin (BCG) has long been approved as an immunotherapeutic agent , tetrahydropyranyl-adriamycin (THP) , gemcitaic agent  and its ic agent .Clinically, intravesical drugs can reduce tumor recurrence rates to some extent. However, the treatment effect is significantly lower than expected, which may be attributed the following factors: 1) the drugs lack selectivity for tumor cells, leading to limitation of the tumor-killing effect, and exert significant toxic effects on normal bladder mucosa ; 2) due Targeted therapy has received considerable research attention for further development. Targeted drug delivery systems present significant advantages in the treatment of bladder cancer by facilitating selective delivery of chemotherapy to tissues. However, achievement of therapeutic drug delivery specifically to tumor cells or tissues through the membrane and subsequent release of the drug into target tumors without damage to normal cells remains a significant challenge . ResearcNanotechnology mainly deals with materials between 1 and 200 nm in size . Since tNanoparticles used as chemotherapy drug carriers are submicron-sized particles (3–200 nm), devices or systems generated using different materials including polymers, such as micelles, dendrimers, liposomes, viral nanoparticles, and even organometallic compounds . The nanThe efficacy of conventional instilled chemotherapy for bladder cancer is adversely affected by limitations in the ability of drugs to reach tumors . During Nanocarriers are commonly used to enhance the targeted release of drugs  but can Several types of nanoparticles have been investigated in clinical medicine applications including disease diagnosis and treatment. The carrier function of nanoparticles for drug delivery is a considerable focus of research. All nanoparticle applications are based on the uptake and utilization of nanomedicines by cells, with surface charge determined as an important factor affecting cellular uptake . AccordiChitosan (polyglucosamine (1–4)-2-amino-BD glucose) is a product of deacetylation of chitin. Chitosan is the only polycationic polysaccharide in nature with unique biological properties amenable for clinical use, such as non-toxicity, good biocompatibility, and easy degradation . This liThe inner bladder surface has a biological barrier that prevents drug penetration. The bladder permeability barrier is caused by the thick mucus gel layer coating the bladder mucosa aligned with glycosaminoglycan chains dispersed in a viscous hydrogel .2O2 in the targeted tumor sites to generate O2 through catalase and improved the therapeutic effect of SDT in ablation of bladder cancer under ultrasound  mucosal platform that autonomously generates transmucosal oxygen. In this system, fluorinated chitosan was used as an efficient and non-toxic transmucosal delivery carrier loaded with meso-tetra(4-carboxyphenyl) porphine-conjugated catalase. CAT-TCPP/FCS nanoparticles (NP) instilled into the bladder showed excellent transmucosal and tumor penetration ability. Simultaneously, CAT-TCPP/FCS nanoparticles effectively alleviated hypoxia in the tumor by catalyzing Htrasound   was used to load the intravesical chemotherapeutic drug mitomycin C (MMC). Notably, CS-PLC nanocarriers remained in the bladder for a long time, accumulated in bladder tumors, and prevented drug loss during urine excretion .L-lysine (PLL), are commonly used as a positively charged drug delivery vehicle  . As showCompared with the traditional method of administration, namely, direct intravesical drug instillation therapy, nanocarriers or biological materials exert greater effects on eliminating tumors and inhibiting tumor growth. However, ordinary nanomaterials have a number of drawbacks . The priFolic acid  is a critical nutrient for growth and development of organisms that plays an indispensable role in DNA methylation, biosynthesis and repair. In nature, folic acid is widely distributed in various fruits and vegetables . Two mec4. Next, the photosensitizer methylene blue (MB) was combined on the surface of Au@TNA to generate Au @TNA@MB nanoparticles. Subsequently, nanocarriers were modified with folic acid (FA) to obtain FA-Au@TNA/MB NPs. The newly generated nanocarrier was composed of two types of nanoparticles  with an average size of 26.43 ± 3.7 nm -assisted reduction of HAuCl± 3.7 nm . In vitrSV-HUC-1  but aggrSV-HUC-1 . TherefoSV-HUC-1 .Compared with other targeted therapy, peptide-modified nanocarriers offer several advantages including lower production costs, relative stability, effortless commercialization, simple operation, and reduction of the immune response. At the same time, binding of peptides to receptors can achieve precise targeted therapy through design of specific reactive groups. In addition, particle size is relatively low, which has little effect on the physical and chemical properties of the synthesized nanoparticles .10 known peptides exist in the phage library  showed strong selective binding to bladder cancer cells. Fluorescent labeling experiments further revealed that CSNRDARRC co-localized with cytokeratin in bladder cancer tissues. Importantly, the peptide was not detected in normal bladder tissues and other organs. The tumor cell selectivity of CSNRDARRC and its specific targeting of bladder cancer strongly support the utility of this peptide as a BC-targeted therapeutic agent .Jung et al. utilized the CSNRDARRC peptide (Bld-1) as a targeting ligand to selectively deliver KLA to bladder cancer and examined the activity of Bld-1-KLA hybrid peptide as targeted therapy for bladder tumors with pro-apoptotic peptides. The hybrid peptide effectively targeted bladder cancer cells and induced apoptosis, and could therefore serve as an excellent bladder tumor-targeting pro-apoptotic peptide .In addition, the Bld-1 peptide itself may be applied as a targeting ligand in combination with nanocarriers loaded with conventional chemotherapeutics for drug delivery to bladder tumors. Wei et al.  describeIn vivo targeting of human primary bladder cancer cells was a subsequent focus of a study by Zhang et al. A one-bead one-compound combinatorial peptide library technology (OBOC) was applied to identify several bladder-specific ligands with the same DGR motif showing potential utility in targeted bladder cancer therapy. However, only the cyclic PLZ4 peptide with the amino acid sequence cQDGRMGFc was specific for bladder cancer. PLZ4 could specifically bind αvβ3 integrin on BC cells but not normal urothelial cells  has multiple applications in photodynamic diagnosis, photothermal therapy and targeted chemotherapy. PNP is a nanosphere that emits fluorescence/heat/reactive oxygen species upon irradiation with near-infrared. Compared with free Dox, the PLZ4-PNP platform significantly prolonged overall survival of mice. The three effective modes  of the platform could be easily applied for individualized treatment and have significant potential in the targeted management of bladder cancer .in vivo and in vitro. As shown in Pan and co-workers developed bladder cancer-specific nanomicelles loaded with the chemotherapeutic drug paclitaxel (PTX). The micelles were gradually combined with polyethylene glycol, cholic acid and PLZ4. PTX and micelles were mixed and encased in the core, finally forming disulfide-crosslinked PLZ4 nanomicelles (DC-PNM-PTX). The drug loading rate of PTX was 25% and drug loading efficiency was >99%. The nanomicelles were spherical with a particle size of about 25 nm. Notably, upon intravenous administration in vivo, DC-PNM-PTX targeted bladder cancer but had no specificity for lung cancer in the same animal (The group of Lin further combined one end of polyethylene glycol with a cholic acid cluster and modified the other end with PLZ4. Daunorubicin (DNR) and paclitaxel (PTX) were loaded into the prepared micelles to form PLZ4-modified PLZ4-DNR/PTX nanoparticles with a particle size of about 23.2 ± 8.1 nm and their efficacy in dog bladder cancer examined e animal .PLZ4 has therefore been identified as a target peptide for bladder cancer and effectively utilized as a modified vehicle for accurate delivery of drugs to achieve the maximum therapeutic effect.Polybia paulista  peptide sequence with remarkable targeting ability specifically interacts with cells overexpressing αvβ3 integrin and plays a critical regulatory role in tumor growth, metastasis and angiogenesis . High-af1H-NMR analysis and affinity for human glioblastoma-astrocytoma cells (U87MG). The in vitro cytotoxicity assay was used to evaluate the viability of bladder cancer cells (T-24) after incubation with doxorubicin (DOX) polymer micelles. The results showed strong affinity of c (RGDfK)-modified micelles for T-24 cells and significant inhibitory effect on cell proliferation when loaded with doxorubicin drugs, supporting a significant effect of c (RGDfK) on bladder cancer. The c (RGDfK)-modified micelles assembled by PCL-b-PEO diblock copolymer developed in this study displayed remarkable potential as a nano-drug system for superficial bladder cancer bladder perfusion chemotherapy  is a focus of increasing research attention in the field of cancer treatment. EGFR is a member of the tyrosine kinase receptor family and its abnormal expression is associated with excessive cell proliferation. In addition, EGFR is reported to promote angiogenesis in tumors and inhibit growth and development of tumor cells . EGFR isIn terms of nanotherapy, Cho and co-workers synthesized multifunctional nanoclusters of upconversion nanoparticles (UCNP) and gold nanorod (AuNR) through PEGylation. The UCNP-AuNR multifunctional nanoclusters generated could be effectively applied to treat bladder cancer. Subsequently, these nanoclusters were further modified with antibodies to generate functionalized UCNP-AuNR with C225 antibody. In cell experiments, functionalized nanoclusters exerted obvious cytotoxic effects relative to those without antibody modifications. The use of monoclonal antibodies to modify nanocarriers is proposed to effectively reduce the dosage and maximize the therapeutic effects of drugs .Owing to its multiple biological properties, such as high hydrophilicity and swelling capacity, hyaluronic acid (HA) is utilized in various drug carrier systems . HA is ein vivo and in vitro experiments showed that siRNA@CS-HAD nanoparticles effectively suppressed bladder cancer growth without exerting significant biological toxicity. The delivery of cy3-siRNA promoted targeted gene silencing. In vivo, siRNA@CS-HAD NPs accumulated in BC tissues and exerted a strong inhibitory effect on target oncogenes and tumor growth. This novel nanosystem may therefore present an effective method for targeted treatment of BC with high CD44 expression (Liang and co-workers confirmed markedly higher expression of CD44 in bladder cancer relative to normal bladder. CD44 was further identified as a specific receptor for HA. Combination of HA and CD44 could induce conformational changes and allow adaptor proteins or cytoskeleton elements to bind intracellular domains, thereby activating multiple signaling pathways for cell proliferation, adhesion and metastasis . The autpression  (Figure pression .Bladder cancer is a global disease with high incidence worldwide. Treatment of bladder cancer generally entails surgical removal followed by intravesical instillation of chemotherapy into the bladder to prevent recurrence. However, conventional chemotherapeutic drugs are not selective for normal tissue cells and trigger numerous side-effects when frequently instilled. For example, instillation of conventional chemotherapeutics into the bladder is associated with chemical cystitis and hematuria, with the severity being significantly dependent on the dose and frequency of instillation. Moreover, the majority of adverse effects can be improved after termination of drug treatment. Due to the higher recurrence and progression rate of BC, patients require long-term and repeated intravesical instillation after surgery. However, frequent intravesical instillation of chemotherapy is associated with several problems, such as high cost and excessive family burden. In recent years, nanotechnology has been applied to resolve these issues and shown to be feasible and efficient for diagnosis and therapy of bladder cancer.According to their characteristics, nanoparticles are divided into actively targeted and passively targeted therapeutic carriers. The fundamental principle of passive targeted therapy is the EPR effect while the active targeted drug delivery system uses specific ligands for attachment to vehicles or modification of the nanocarrier surface. Using the above mechanisms, nanoparticles can overcome the limitations of current traditional drug treatments for tumors, leading to significant improvement of therapeutic activity along with reduction of the side-effects of drugs. For instance, the nanoparticle surface induces significant changes in nanocarrier properties and interactions with the surrounding environment. At the same time, nanoparticles carry charges and non-specifically bind tumors through electrostatic interactions. As described above, positively charged nano-drug delivery carriers formed by chitosan bind non-specifically to the negatively charged inner wall of the bladder, leading to enhanced drug pairing. Adhesion to the bladder wall prolongs contact time with the tumor and enhances drug effects to an extent. Increasing attention has focused on tumor-specific targeted therapy and further research is ongoing.in vivo and in vitro experiments on nanoparticles modified with targeting ligands and encapsulating chemotherapeutic drugs have confirmed that receptor-mediated endocytosis increases chemotherapeutic drug uptake by cancer cells. Targeted nanoparticle delivery carriers have several advantages over general nanocarriers. Firstly, these carriers are selective for bladder cancer cells, reducing their combination with normal bladder tissues and cells, and decreasing the side-effects caused by drug perfusion in the bladder. Secondly, drug-loaded nano-systems accumulate at tumor targets and release the drug, increasing therapeutic drug concentrations at the tumor tissue sites. Thirdly, specific ligand-receptor binding can extend the drug retention time in the bladder and prevent its excretion with urine. Fourthly, the release time and speed of the drug can be controlled to reduce or even avoid irritation to the bladder caused by multiple infusions of the drug.Targeting ligands bound to the surface of nanoparticles can efficiently recognize receptors expressed on cancer cells, significantly improving the selectivity and killing effects of chemotherapeutics on bladder cancer cells. Targeting ligands generally include protein/peptide, hyaluronic acid, folic acid, and polysaccharides. Numerous Despite the numerous benefits of targeted nanoparticles in tumor treatment, several problems need to be resolved before clinical application, for example, between the experimental stage of nanomedicine and its transformation into a clinical agent, the technical problems of large-scale production of nanomedicines, and cost control during the manufacturing process. Moreover, immunotherapy has also made major clinical breakthroughs and it may be applied to bladder cancer in the future, which will greatly improve the therapeutic effect of bladder cancer .In summary, although the efficacy of nanotherapy for bladder cancer remains to be established, its potential in targeted treatment and drug release provides a sound theoretical and practical basis for application in personalized treatment of bladder cancer.


Approximately 1 in 8 men will be diagnosed with prostate cancer (PCa) at some point in their lifetime. Five-year survival rate for patients with local or regionally spread disease is near 100%, but this drops to 30% when the cancer has spread to distant sites. Androgen receptor (AR) signaling plays a pivotal role in normal prostate development and PCa cell survival. Therapies targeting the AR pathway are mainline treatments, but resistance is a major clinical problem. From the literature and our own meta-analysis of PCa databases, we aimed to review the connections between PCa genetic alterations and the AR signaling axis at the primary stage. Assessing how combinations of PCa genetic drivers that arise in patients affect AR signaling will aid in stratifying patients who will likely respond to AR-directed therapies, and those who will require other therapeutic agents upfront in order to prevent disease progression.AR gene itself occur at later stages, genomic changes at the primary stage can affect the AR axis and impact response to AR-directed therapies. Here, we review common genomic alterations in primary PCa and their influence on AR function and activity. Through a meta-analysis of multiple independent primary PCa databases, we also identified subtypes of significantly co-occurring alterations and examined their combinatorial effects on the AR axis. Further, we discussed the subsequent implications for response to AR-targeted therapies and other treatments. We identified multiple primary PCa genomic subtypes, and given their differing effects on AR activity, patient tumor genetics may be an important stratifying factor for AR therapy resistance. While many prostate cancer (PCa) cases remain indolent and treatable, others are aggressive and progress to the metastatic stage where there are limited curative therapies. Androgen receptor (AR) signaling remains an important pathway for proliferative and survival programs in PCa, making disruption of AR signaling a viable therapy option. However, most patients develop resistance to AR-targeted therapies or inherently never respond. The field has turned to PCa genomics to aid in stratifying high risk patients, and to better understand the mechanisms driving aggressive PCa and therapy resistance. While alterations to the  Prostate cancer (PCa), the most commonly diagnosed cancer in men in the United States, remains the second leading cause of U.S. male cancer related deaths. The majority of cases (89%) are diagnosed when the disease is localized to the prostate and neighboring organs, providing a five-year survival rate of nearly 100%. However, once the disease has spread to distant sites, patients have a significantly lower five-year survival rate of 30% . Lo. LoPTEN, outcome ,69. Althectively .FKBP5, stabilizes PHLPP1 and PHLPP2, important phosphatases in the pathway, allowing for deactivation of phospho-AKT. This feedback loop can, in turn, contribute to AKT-driven growth in castrate conditions, as androgen deprivation leads to decreased FKBP5, PHLPP1 and PHLPP2 and increased AKT activation. AKT is also able to phosphorylate AR and mark it for ubiquitination and degradation [PTEN loss displayed significantly below average AR scores in multiple cohorts , PARP1 (8–10%), BRCA1 (4–11%) and BRCA2 (6–22%) are some of the most common DNA repair genes deleted or mutated in primary PCa (BRCA1/2 can result in greater dependency on error-prone non-homologous end joining (NHEJ), consequently increasing the mutational burden in PCa [PARP1 and ATM also contribute to the genomic instability of PCa through their critical roles in recruiting repair proteins and coordinating repair with cell cycle arrest, respectively [Many genes involved in DNA repair are altered in PCa, and this in turn contributes to the genomic instability and array of genomic alterations present in PCa. mary PCa . Changesn in PCa . The losectively ,97.PARP1 loss to decrease AR activity in patients. While PARP1 deletions are associated with average AR mRNA levels, they are consistently associated with above average AR scores in multiple cohorts  are promising therapeutic strategies under investigation ,135,136.There are however a few caveats to our own analysis, as well as significant hurdles when it comes to the application of tumor genomics in the clinic. The currently available patient data for PCa, including the cohorts used in this review, largely involve bulk tumor RNA-Seq. This along with tumor heterogeneity limits some of the interpretations that can be made about genomic alterations in patients. It is unclear if some of the co-occurring alterations occur in the same cells, mosaically within the same tumors, or represent independent multifocal areas of disease. The multifocal and heterogeneous nature of PCa poses challenges for applying tumor genomic analysis to inform treatments. The growing application of single cell -omics analysis will hopefully aid in elucidating the nature of co-occurring alterations, as well as their effects on the AR axis. With continued advancements in the field and more clinical studies, tumor genetics could be used to inform treatment decisions. This may help delineate patients who will likely do well on AR monotherapies, be inherently resistant to AR therapies, or need treatment combinations to combat the inevitable acquired resistance to AR single agent therapies.


Obesity is a significant risk factor for various cancers including breast cancer resulting in an increased risk of recurrence as well as morbidity and mortality. Extensive studies on various pathways have been successful in establishing a biological relationship between obesity and breast cancer. The molecular classification of breast cancer includes five groups each having different responses to treatment. Increased levels of inflammatory cytokines seen in obese conditions drive the pro-proliferative pathways, such as the influx of macrophages, angiogenesis, and antiapoptotic pathways. Increased peripheral aromatization of androgens by aromatase increases the circulating estrogen levels which are also responsible for the association of obesity with breast cancer. Also, increased oxidative stress due to chronic low-grade inflammation in obese women plays an important role in carcinogenesis. Despite the availability of safe and effective treatment options for breast cancer, obese women are at increased risk of adverse outcomes including treatment-related toxicities. In the recent decade, selenium compounds have gained substantial interest as chemopreventive and anticancer agents. The chemical derivatives of selenium include inorganic and organic compounds that exhibit pro-oxidant properties and alter cellular redox homeostasis. They target more than one metabolic pathway by thiol modifications, induction of reactive oxygen species, and chromatin modifications to exert their chemopreventive and anticancer activities. The primary functional effectors of selenium that play a significant role in human homeostasis are selenoproteins like glutathione peroxidase, thioredoxin reductase, iodothyronine deiodinases, and selenoprotein P. Selenoproteins play a significant role in adipose tissue physiology by modulating preadipocyte proliferation and adipogenic differentiation. They correlate negatively with body mass index resulting in increased oxidative stress that may lead to carcinogenesis in obese individuals. Methylseleninic acid effectively suppresses aromatase activation thus reducing the estrogen levels and acting as a breast cancer chemopreventive agent. Adipose-derived inflammatory mediators influence the selenium metabolites and affect the proliferation and metastatic properties of cancer cells. Recently selenium nanoparticles have shown potent anticancer activity which may lead to a major breakthrough in the management of cancers caused due to multiple pathways. In this review, we discuss the possible role of selenoproteins as chemopreventive and an anticancer agent in obese breast cancer. Glandular tissues include the milk-producing glands (lobules) and ducts (milk passages), while stromal tissues are made up of fibrous and fatty connective tissues of the breast. Cell fluids and waste are extracted by the breast lymphatic tissue immune system. Microarray-based gene expression profiling has revealed that breast cancers are intricate, and their histology is highly variable. Nevertheless, these studies have contributed their experimental results to classify tumors into different types based on the lymph node status, tumor grade, molecular type, and the presence of predictive markers such as estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor (HER2) . A prognsubtypes . Cancer subtypes . The extsubtypes . Invasivsubtypes . These csubtypes .in-situ (DCIS) is the most common non-invasive breast cancer  is non-invasive breast cancer and is considered as one of the markers to predict the increased breast cancer risk (in situ (PLCIS) is a variant of LCIS and shows similar histopathological features as DCIS. PLCIS is further classified into 1) non-apocrine and 2) apocrine  remodeling. Reciprocal interactions between epithelial cells and adipocytes are often identified during the cancer process: tumor development, tumor growth, invasion, and metastases . SeveralLike leptin, insulin is also a major risk factor for the development of breast cancer in postmenopausal women as it induces the activity of adipose aromatase. In a more complex network between leptin and other factors, leptin and estrogen metabolism are shown to be related. Consequently, obesity-associated hyperleptinemia is suggested as a primary mediator for breast cancer pathophysiology . Higher Leptin has also been shown to facilitate the growth and progression of neoplastic breast cancer cells by stimulating the pathway of Janus Kinase/Signal transducer and activator of transcription (JAK2/STAT3). Once leptin attaches to its receptor, the leptin receptor cytoplasmic domain recruits JAK2 kinase and phosphorylates the tyrosine residue. This results in the STAT3 protein anchorage, which is recruited from the SH2 domain. When STAT3 is bound to the receptor, the proteins JAK2 and STAT3 are phosphorylated to dimerize, allowing them to be translocated to the nucleus. They act as transcription activators for a variety of genes, such as c-myc, cyclin D1, p21/waf1, c-jun, junB, erg-1, and Bcl-2, all of which are involved in cell growth and proliferation . InsulinObesity being a state of chronic low-grade sterile inflammation presents with increased levels of circulating pro-inflammatory factors without clinical signs of inflammation . AdiposeP (SePP1) are the important selenoproteins present in our system.Enzymes such as glutathione peroxidase (GPx), thioredoxin reductases (TR), iodothyronine deiodinases (DIO), selenoprotein In-vitro and in-vivo studies in recent years have come up with some interesting results where GPx7 acts as a catalyst for oxidative protein folding by utilizing Ero1-flavoproteins generated hydrogen peroxide (H2O2) suggesting GPx7 as a scavenger for hydrogen peroxide . They are the central enzymes in the Trx redox pathway. Trx acts as a reducing substrate in the reduction of ribonucleotides and detoxification of radicals and oxidants . TrxR faectively . Sec is ectively . Trx/Trxectively . Inhibitectively . Thioredectively . InhibitDIO is a family of selenoenzymes that consists of three members, DIO1, DIO2, DIO3, and share catalytic and sequences homology . DIOs arSepp1 is a secreted glycoprotein found in plasma containing multiple Sec residues and thought to have antioxidant properties. Selenium is regulated in the liver and is transferred to the organs utilizing Sepp1 which serves as a measure of selenium intake status . The selß-oxidation, Se suppresses the formation of fatty liver and induces inhibition of adipocyte hypertrophy and abdominal fat accumulation  converts androgens to estrogens and is a key enzyme for the biosynthesis of estrogen. Hence, aromatase is an important target for breast cancer prevention and therapy as estrogen plays a critical role in obese postmenopausal breast cancer progression. Nevertheless, treatment with aromatase inhibitors is associated with several adverse effects like osteoporosis, cardiovascular, and neurocognitive defects due to systemic suppression of estrogen biosynthesis . MethylsPyracantha fortuneana has shown to have anticancer activity against TNBC MDA-MB-231 cells by arresting the cells in the G2 phase and inhibiting CDC25C-CyclinB1/CDC2 pathway (ob/ob) mice wherein complex-I is identified as a critical modulator of obesity-induced metabolic remodeling in ß-cells which in turn is regulated by cyclin-dependent kinase-1 (CDK-1) (in-vitro (Triple-negative breast cancer (TNBC) is an aggressive type of breast cancer that does not respond well to hormone therapies. Treatment options for TNBC are very limited and immediate needs for alternative therapies are indeed in demand. There are several mechanisms linking obesity and TNBC, including the role of insulin on the AKT/mammalian target of rapamycin (mTOR) signaling network, tissue inflammation, and protumorigenic tissue microenvironment driving the aggressive TNBC . Seleniu pathway . This pa (CDK-1) . It has  (CDK-1) . Selenofin-vitro . The att. GPx3 has been identified as a regulator of insulin receptor (IR) expression and insulin sensitivity in adipose tissue , GPx1 decreases the cell adhesion and proliferation as well as decreases the metastasis of TNBC cells to the lungs . A recene tissue . Downreg+Ly6G+ neutrophils in an ovariectomized C57BL/6 J female mice. Hence, they concluded that the incidence of metastasis and a higher ratio of triple-negative breast cancer is seen in obesity  intravasation of macrophages via the epidermal growth factor or colony-stimulating factor 1, 2) signaling pathway of paracrine, 3) the microenvironment of the tumor, 4) the vessel wall elements surrounding the intravasation of tumor cell clusters, 5) cooperative intravasation and 6) intravasation associated with the vasculogenic mimicry . In 40% The selenoproteins show a pivotal role as an anti-metastatic drug by involving in redox reactions, homeostasis of calcium, and mitigation of stress . In a stLactobacillus bravis enriched with selenium nanoparticles (SeNPs) increased the lifespan of tumor-bearing BALB/c mice by a significant elevation in serum IL-17 and interferon γ (IFN-γ) and natural killer (NK) cell activity and also reduces liver metastasis  and redox enzymes and also reducing oxidative damage to proteins and lipids. In contrast to this, selenocysteine decreased the antioxidant enzyme and UCP2 protein expression, thereby increasing the ROS production and decreasing the viability of breast cancer cells . UpregulThe damaged and dysfunctional intracellular components are delivered to the lysosomes for degradation by a process that is referred to as autophagy which is regulated by mTOR signaling. Compounds that induce or inhibit autophagy may be exploited for therapeutic applications against cancer. Seleno-purine (SLNN-15) is shown to stimulate anti-proliferative activity in MDA-MB-231 cell lines via inducing autophagy by selectively inhibiting the AKT-mTOR pathway . UpregulMany cancer trials indicate that glutathione is of major significance in breast cancer. It is established that Glutathione depletion makes cancer cells vulnerable to chemotherapeutic drugs. Other effects of glutathione depletion are shown in Selenoproteins are potent modifiers of carcinogenesis and tumor progression. They inhibit tumor development by revoking the oxidative stress insults particularly in cancers that are driven by inflammatory mediators. Breast cancer and obesity both are chronic inflammatory conditions that present with increased oxidative stress and altered redox homeostasis. Decreased selenoproteins in adipose tissue may result in adipocyte dysfunction leading to insulin resistance, inflammation that may progress to breast cancer. Selenium supplementation may help in mitigating oxidative stress thus lowering the occurrence of carcinoma. However, this has been an area of debate as selenium supplementation without assessing the previous selenium status in individuals might be hazardous. Also, further studies have to be undertaken to precisely understand the biological mechanism of how selenium influences the tumorigenesis in obese breast cancer that belong to different molecular subtypes.


With the advent of newer technologies, real-time anterior segment optical coherence tomography (OCT) integrated with the operating microscope has become possible. We are proposing the technique of bleb revision with greater precision under direct visualization of bleb anatomy and extent of tissue depth allowing better localization and easy maneuvering with lesser complications. In this surgical technique, bleb revision was performed using intraoperative real-time OCT incorporated in OPMI LUMERA 700 microscope. Live surgical and OCT view were seen on a common screen together. A moderately elevated, diffuse functional bleb was noted after three months of bleb revision in both cases with controlled intraocular pressure. Intraoperative OCT-guided bleb-sparing epithelial exchange is an adjunctive technique for bleb repair surgery with an increased precision of surgery which can reduce complications, minimize surgical time and maximize surgical outcome. The intraoperative use of MMC and 5-fluorouracil in trabeculectomy has increased the susceptibility of thin avascular blebs. These thin avascular blebs are more prone to vision-threatening complications such as hypotonic maculopathy, blebitis, and endophthalmitis, especially in pediatric age groups. Ehlers et al stated that the use of microscope-mounted intraoperative optical coherence tomography (iOCT) can modify the surgical decisions of both anterior and posterior segment surgeries.,4 Various surgical or nonincisional methods using cyanoacrylate glue or autologous blood injections for bleb revision have been described.,6,7 Our technique of bleb-sparing epithelial exchange (BSEX) is well established and safe.,9Long-term results of trabeculectomy have improved over time with modifications to surgical technique, However, the trypan blue dye used for staining the epithelium causes difficulty in visualizing the epithelium and thereby dissection. This may either lead to incomplete epithelium removal or damage to the underlying bleb structure. Peeling in multiple fragments is required in these cases to avoid inadvertent damage to the underlying bleb structure, which might not ensure complete removal. Therefore, we used iOCT in adjunct to trypan blue dye to ensure complete epithelial peeling without causing any inadvertent damage to the underlying functional bleb structure.The use of iOCT is a noninvasive tool evolving for different glaucoma surgical interventions like trabeculectomy, goniosynechialysis, Ahmed glaucoma valve (AGV) implantation, bleb needling, etc. In this technique, epithelial peeling of a thin avascular bleb followed by conjunctival advancement is done, which preserves the functional bleb without excising it and thereby less need of glaucoma medications postoperatively.Bleb revision was done after taking a written informed consent, using an iOCT RESCAN 700  fused with OPMI LUMERA 700 microscope . The Calisto eye displays the live surgical and OCT view on the common screen together.Case 1: A 58-year-old male with primary open-angle glaucoma presented with a past history of right eye (RE) trabeculectomy seven years back. On examination, RE vision was 6/9 on Snellen's visual acuity chart and IOP was 8 mmHg. Slit lamp examination showed a thin avascular bleb with sweating on fluorescein staining. [Figures 1a and 1b]. Bleb anterior segment optical coherence tomography (ASOCT) showed an overhanging and elevated bleb [Figure 1c]. RE iOCT-guided bleb revision was performed.Case 2: A 36-year-old female with juvenile open-angle glaucoma presented with left eye (LE) IOP of 4 mmHg. She had a history of both eyes trabeculectomy done one year ago. On slit lamp examination, a thin avascular bleb with a bleb leak was present. She also complaint of drop in vision of LE and her visual acuity on presentation was 1/60, with evidence of hypotonic maculopathy visible in fundus. She underwent LE iOCT-guided bleb revision.In vivo, preoperative and postoperative assessment of the bleb was done on iOCT [Figures 2a and 2c]. Postoperatively, topical antibiotic–steroid combination eyedrops and ointment were prescribed for four weeks. Both patients were reviewed at day 1, week 1, month 1, and month 3 after the surgery.After anesthetizing the eye with peribulbar block (0.5% bupivacaine + 2% lignocaine), a wire speculum was inserted. A curved Vanna's scissor was used to cut the conjunctiva along the lateral and the posterior margins of the avascular bleb. Hemostasis was achieved and the surrounding normal conjunctiva was separated from the underlying Tenon's with a blunt-tipped Westcott's scissor, until its free margin could be advanced easily to the limbus. Care was taken to avoid buttonholing of the conjunctiva. Trypan blue dye (0.06%) was applied to stain the epithelium over the bleb. The anterior edge of the bleb epithelium was identiﬁed, and only the stained epithelium was gradually peeled off the surface backward and horizontally under direct visualization of the tissue depth on iOCT. This prevented damage to the underlying functional bleb structure [Figure 2b]. As the raised epithelial sheet reached the more ﬁbrosed posterior margin of the bleb, it was cut off. Thereafter, the adjacent conjunctiva was advanced and hitched to the limbus with 6-0 vicryl horizontal mattress sutures. A watertight closure of the conjunctival ﬂap was achieved by continuous 8-0 vicryl sutures at the lateral edges avoiding dog ears and two 10-0 nylon horizontal mattress sutures, buried across the limbal extent. The first case after bleb revision showed an RE diffuse and vascular bleb with an IOP of 14 mmHg on postoperative day 1. RE vision was maintained at preoperative levels at three months follow-up. Bleb sweating was absent. A moderately elevated, diffuse functional bleb was noted after three months of bleb revision with RE IOP of 12 mmHg. The ASOCT of the ﬁltering blebs postoperatively showed a decrease in the bleb height, a thicker epithelium, and smaller hyporeﬂective spaces within the bleb tissue [Figures 3a and 3b]. The second patient also showed a functional bleb at four months postoperatively. The vision in LE improved to 6/24 with resolution of hypotonic maculopathy. Ever since, its use in ophthalmology has evolved. iOCT has been used for bleb needling for an encysted bleb with high IOP after trabeculectomy, important guide during crucial steps of lamellar keratoplasty, as well as during stromal dissection in ocular surface squamous neoplasia.,11Introduction of OCT into clinical practice brought a landmark change in diagnosis, management, and monitoring of nearly all ocular pathologies. Along with that, OCT integrated with operating microscope resulted in a paradigm shift in ocular surgery. The feasibility of iOCT was first described by Ehlers et al.During surgical maneuvering, the iOCT with heads up display (HUD) allows rapid visualization of the area of interest and provides the surgeon with information regarding instrument and tissue interactions. Because of its finer resolution, OCT is able to present detailed view of the very thin bleb wall, and an accurate assessment of separating epithelium from functioning bleb structure.The epithelium can be precisely identified and dissected under direct visualization without causing any inadvertent damage to the underlying bleb structure. This permits better maneuvering, improved dissection of the anterior and posterior bleb wall under visualization, thereby reducing the risk of complications. Moreover, iOCT may also be useful in visualizing any underlying scleral defect. However, repeated microscope adjustments for a better focus of the area of interest and obscuration of the operation site due to conjunctival bleeding may cause difficulties during the procedure.,12 It also reduces the direct trauma to the underlying bleb structure and can also be used as an excellent teaching tool for trainees. However, the high cost and availability of iOCT are the major limitations. This technique could be applied to a wider range of patients including all age groups. In summary, iOCT-guided bleb-sparing epithelial exchange is an adjunctive technique for bleb repair surgery which can maximize the surgical outcome, minimize surgical time, and reduce complications with an increased precision of surgery. Further studies with a larger sample size and a longer follow-up along with a comparison to standard procedures are recommended.iOCT ensures complete epithelial peeling of the deceased epithelium, which is important to prevent postoperative increase in IOP.Nil.There are no conflicts of interest.


Glioblastoma is an aggressive tumor and is associated with a dismal prognosis. The availability of few active treatments as well as the inexorable recurrence after surgery are important hallmarks of the disease. The biological behavior of glioblastoma tumor cells reveals a very complex pattern of genomic alterations and is partially responsible for the clinical aggressiveness of this tumor. It has been observed that glioblastoma cells can recruit, manipulate and use other cells including neurons, glial cells, immune cells, and endothelial/stromal cells. The final result of this process is a very tangled net of interactions promoting glioblastoma growth and progression. Nonetheless, recent data are suggesting that the microenvironment can also be a niche in which glioblastoma cells can differentiate into glial cells losing their tumoral phenotype. Here we summarize the known interactions between micro-environment and glioblastoma cells highlighting possible therapeutic implications. The 2021 World Health Organization (WHO) Classification defines glioblastoma (GBM) as a diffuse astrocytic glioma without IDH (Isocitrate dehydrogenase) and H3R gene mutations, with enhanced microvascular proliferation, necrosis, and specific alterations like gain of chromosome 7/chromosome 10 loss, EGFR  amplification, and/or TERT (Telomerase Reverse Transcriptase) mutations  malignancies with an estimated 5-years overall survival (OS) of only 6.8% , 4. In pSeveral efforts have been spent to improve clinical outcomes of patients with GBM, nonetheless, none of the investigated agents have replaced the standard of care represented by maximal safe surgery followed by chemoradiation and adjuvant temozolomide, which has been adopted in 2005 . Even inThere are several reasons behind these failures. First, the enrollment of GBM patients within clinical trials is limited only to10% of all GBM patients , 8.However, the major cause explaining the difficult development of effective agents is certainly the complex biology of the disease. Indeed, GBM is associated with specific features which can be summarized in 1) the extremely high heterogeneity presented by tumor cells requiring combined treatment for different subtypes of GBM cancer cells; 2) the lack of biological models able to replicate or at least estimate the interaction between human tumor cells and the surrounding tissue; 3) the complex microenvironment surrounding the tumor –12. FurtOf note, it has been demonstrated that GBM cells can manipulate the microenvironment surrounding themselves developing a niche sustaining tumor growth and development . This prOf interest, GBM cells can develop a nuclear and cytoplasmatic ‘‘continuum’’ with neighboring cells. These nanotubes mediate the transfer of protein and inorganic elements . MoreoveGBM cells can also promote the creation of gap junction and cytoplasmatic connections –20. FinaThe interactions between tumor cells and immune cells have acquired an increased interest due to the availability of agents able to promote immune-system reactivation. Nonetheless, other interactions have been only partially investigated and could hide novel promising targets for tumor treatment. Here we performed a review summarizing the identified interactions between GBM and its associated microenvironment. We also focused our attention on possible novel treatments targeting these complex interactions.Immune-contexture assumes a very important role within the GBM microenvironment. It can stimulate the progression and development of tumor cells .In contrast to what was supposed in the past, it has been largely demonstrated that glioblastoma is not an immune ‘‘cold tumor’’ , 27. SimRecently, two transcription factors showed to mediate several immune-depressive effects in GBM . These fAnother recent research investigated RNA expression of GBM-derived sphere-line treated with the BET inhibitor JQ1. The inhibition of BET resulted in a significant modulation of genes responding to Interferon-alpha (enhanced in about 50% of GBM) through a direct transcriptional inhibition more than interference to the JAK (Janus Kinase) -STAT  pathway .BET inhibitors have recently been assessed on phase I clinical studies and further trials assessing their clinical efficacy are needed –47.In conclusion, differently from other tissues, CNS physiologically employs signals able to reduce an immune innate and adaptive response. This happened to prevent catastrophic effects associated with an uncontrolled inflammation in a dedicated system such as the brain or the spinal cord. In this context, GBM cells adopt several mechanisms to reinforce this inhibition. Glioblastoma stem cells (GSCs) are probably an important component of this complex mechanism as they express important transcription factors such as Sox2 and Oct4 . NonetheImmune resident cells of the CNS are microglia and macrophages including perivascular, meningeal, choroid plexus, and circumventricular macrophages . These cMyeloid cells constitute a large part of the immune-contexture of GBM. These cells are recruited directly from tumors cells and then can differentiate toward macrophages and monocyte phenotypes .Myeloid cells have been classified into monocytic and granulocytic subtypes. Both these cells inhibit T cell and NK activities. Curiously, some data seem to confirm a negative prognostic role of these cells in males while these same cells can positively activate an immune response in female patients .The main stimulation for macrophages and microglia accrual around the tumor is mediated by the same glioblastoma cells through the production of CCL2 (C-C Motif Chemokine Ligand 2), CCL7 (C-C Motif Chemokine Ligand 7), GDNF , SDF1 , TNF (tumor necrosis factor), VEGF , ATP (adenosine triphosphate), CSF-1 (Colony-stimulating factor 1), GM-CSF (Granulocyte-Macrophage Colony-Stimulating Factor), and expression of OLIG2 (Oligodendrocyte transcription factor 2) , 51, 53.The differentiation of macrophages ranges from two specific phenotypes which are: M1 (immune-response enhancer) or M2 (immune-regulator inhibiting immune response) , 56. AltIt has been demonstrated that macrophages promote GBM growth and progression in different ways .First, the macrophage tumor suppression phenotype inhibits the response of other immune cells surrounding the tumor. This immune inhibition is mainly explicated by myeloid cells and differentiated macrophages which miss the activation of natural killer (NK) cells, the production of interferon γ (INF γ) and TGFβ . These sThe remodeling of ECM is essential for tumor migration and is mediated by the production of inactive metalloproteases enzymes by GBM cells , 58. TheThe administration of macrophages able to restore immune response would be a promising strategy able to partially restore immune-response against the tumor. The development of engineered macrophages (car-M) is at an early stage of assessment but it could be a promising strategy for GBM treatment .Similar to macrophages and microglia also neutrophils and other innate immune cells are attracted by the tumor. In particular, inflammatory factors secreted after a surgical intervention such as IL-8, TNF, and CCL2 can promote neutrophil infiltration in addition to SDF1 and plasminogen activator inhibitor 1 secreted by tumor cells , 62. TheAntigens exposure can start an adaptive immune response in the CNS like observed in other organs.The presence of lymphocytes surrounding the tumor has been largely reported in GBM where their concentration correlates positively with survival , 64.When an antigen is recognized by the immune system, a specific lymphocyte clone targeting the same antigen expands itself starting an adaptive immune response. The antigen-presenting cells (APCs) are essential to start the clonal expansion as these cells mediate the presentation of the antigen to the lymphocytes initiating the immune response , 30. AntEven if lymphocytes can move around GBM mass their ability to start an immune response against tumor is strongly inhibited by several factors. Once lymphocytes come to the peritumoral tissue they are invested by strong immune-depressive signaling mediated tumor cells and GBM associated microenvironment. The TGFβ1 and TGFβ2 molecules are the main characters for this inhibition , 37, 65.It is important to observe that immune-inhibitory signals are largely provided by the microenvironment more than directly by GBM cells. This is a concrete example of how GBM cells manipulate the microenvironment to sustain their growth and expansion . NotablyGBM cells can also release directly inhibitory signals such Fas antigen ligand (FASLG), and other inhibitory molecules. Lymphocytes assume the classical exhaustion phenotype expressing several-inhibitory receptors including the PD-1, T cell membrane protein 3 (TIM3), lymphocyte activation gene 3 protein (LAG3), and T cell immunoreceptor with immunoglobulin and ITIM domains (TIGIT) .As already specified, it is interesting to observe that a large part of inhibitory molecules released by GBM cells can be mediated by the Sox2/Oct4 .The complex network of immune-inhibitory signals can partially explain the failure of clinical trials exploring immune-checkpoint inhibitors (ICIs). To date, three different phases III trials failed to show a clinical benefit from the addition of nivolumab in newly diagnosed and recurrent GBM –70.In the last years, a novel immune strategy employing engineered T-cells (car-T) has been investigated with favorable preliminary results. In GBM the introduction of reprogrammed T-cells could be ineffective due to the immune shield provided by the tumor microenvironment . StrategThe brain is composed of over 60% of the white matter which is largely composed of myelinic axons. Neurons, astrocytes, and oligodendrocytes with other glial cells (including microglia) are the most represented cells in the brain and CNS .The study of the interactions between these components and glioblastoma appears of extreme interest as these are strictly related to some of the most important biological and clinical features of GBM cells.Indeed, connexons between white matter, neurons, and tumors can partially explain the high recurrence rate of the disease as well as the ability to relapse also in a distant site such as the contralateral hemisphere –74. Of iIndeed, the biological niche composed of GBM cells, GSCs, peritumoral oligodendrocytes, astrocytes, and also neuronal axons can both drive differentiation of a GSCs cell toward a tumoral phenotype or, surprisingly, toward a differentiated non-tumoral cell subtype . On the Tumor-associated astrocytes (TAAs) can contribute to GBM growth in different ways. During early phases of tumor development, TAAs respond to initial injury with the secretion of TGFβ, IL-6, and Insulin growth factor 1 (IGF-1) which could contribute to GBM sustainment , 77. TheThe contribution of TAAs is of key importance as these cells can stimulate neo-angiogenesis through secretion of VEGF and hypoxia-inducible factor -1 (HIF-1) , 83.in vitro studies identified that chemoprotection performed by TAAs is possible only when these cells are connected to cancer cells by gap junctions. Indeed, apoptosis and GBM cells death increased when the development of gap junctions was inhibited by the administration of carbenoxolone (a gap junctions communication inhibitor)  .In vivo studies demonstrated that several oncogenes including MYC, RAS, and EGFR variant III can dedifferentiate astrocytes in tumoral cells  can mediate the transformation of astrocyte into malignant glioma cells it is still unclear the exact mechanism by which GBM cells can initiate this process in humans , 86. HowNeurons can be manipulated by glioma cells to stimulate tumor progression and invasion. One of the first documented interactions between neurons and GBM is associated with the release of glutamate which could be enhanced in patients with GBM resulting in seizure onset and increased vascular permeability mediated by the N-methyl-D-aspartate receptors (NMDA) , 91. Of Oligodendrocytes may absolve an inhibitory function against GBM cells. Indeed, these cells can activate the WNT inhibitory factor 1 leading to the inhibition of GBM growth and proliferation , 94. In The mechanism by which this happens involves the white matter destruction which leads to upregulation of the SOX10 transcription factor. The SOX10 mediates oligodendrocyte differentiation also by cancer cells .Further investigation of this pathway will probably provide new therapeutic insights for the clinical management of GBM.Glioblastoma stem cells explain the extreme clinical aggressiveness of GBM. In GSCs are associated with the high recurrence rate of the tumor even after complete resection . These cGlioblastoma stem cells are localized in a specific niche localized in the perivascular and hypoxic regions of the tumor. These cells are similar to normal neural stem cells (NSCs) which are localized mainly in the subventricular zone of the brain which is a common site of origin for glioblastoma –97.It is still unclear if GSCs derived from altered NSCs or mutated glioma cells . GlioblaThe expression of CD133 is regulated by Sox2 and agents targeting or interfering with this transcription factor can reduce tumor-initiating ability, resistance to chemotherapy, and recurrence . Nestin Notably, the specific position of GSCs leads to an interaction with Endothelial cells (ECs) and pericytes. It has been supposed that GSCs directly respond to hypoxia stimulating vessels creation and neo-angiogenesis through the production of VEGF , 104. MoNeovascularization and new vessels development are both hallmarks of GBM. The fast growth of the tumor mass required high blood intake thus there are several identified pathways by which GBM cells can interact and manipulate ECs activity .Hypoxia is one of the most important stimulations for tumor growth, vessels development, and acquisition of more aggressive pathological features by tumor cells –113. In Notably, ROS can assume a different biological role according to their concentration. Low levels of ROS are associated with stimulation of hormone secretion, synaptic plasticity, and immune response , 115. HiOne of the most established pathways for new vessels development is the recruitment of EC progenitors by the bone marrow. This is a process occurring during embryogenesis or after an ischemic insult which is also adopted by GBM cells , 120. TuIt has been well established that GBM cells and the surrounding microenvironment can stimulate the production of the VEGF. This factor can drive EC toward the development of novel vessels in a process known as sprouting angiogenesis. Notably, the neoangiogenic promotion carried out by VEGF can be inhibited by the interaction between the Notch receptor and Delta-like canonical Notch ligand 4 (DLL4) . CuriousVascular co-option is another mechanism by which tumor cells move around pre-existing vessels gaining the access to oxygen and nutrients. Curiously GBM cells can induce secretion of bradykinin by EC cells. Bradykinin plays a chemotaxis effect on GBM cells , 130. ThA well-described mechanism associated with neo-angiogenesis is the interaction between VEGF and VEGFR2 and VEGFR1 . The intThis interaction also results in blood vessels permeability, and proteins lost from blood  explainiThe lack of the survival benefit observed could reflect the co-existence of several mechanisms resulting in angiogenesis and vessels development. In this optic, the inhibition of the VEGF mediated by bevacizumab can activate or reinforce other molecular pathways converging on angiogenesis promotion resulting in resistance to the anti-VEGF.The study of the interactions between immune cells in tumor-associated microenvironment assumes an increased interest . IncreasThus, the inhibition of VEGF could also result in an enhanced immune response against the tumor. This supposed synergic effect has led to the assessment of combination strategies in the clinical setting. Unfortunately, a phase II study assessing the combination between bevacizumab and pembrolizumab (a PD-1 inhibitor) failed to show a significant clinical efficacy on patients with GBM .There are very few systemic agents showing clinical efficacy in patients with glioblastoma –142. As Novel combination strategies are under investigation to overcome these limitations.For example, it has been demonstrated that other immune checkpoints co-exist with PD-1. These are the indoleamine 2,3-dioxygenase (IDO1), T cell immunoglobulin-mucin-domain containing-3 (TIM-3), and lymphocyte activation gene 3 (LAG3) . Novel tPrevious trials investigating neoantigen vaccination showed that vaccines can induce a significant increase of tumors infiltrating lymphocytes . UnfortuSince macrophages are strongly associated with the development of an immune-depressive microenvironment the possibility to develop an engineered cell type of macrophage could be a promising strategy allowing a microenvironment to switch toward an immune-active phenotype . This CaAs already specified SOX2 and Oct4 are two transcription factors able to modulate a very large amount of genes involved in the immune response. These two factors act through proteins belonging to the BET family. To date, BET inhibitors have been assessed on phase I trials with more of them showing a safety profile –47. A fuThe inhibition of the CSF1 receptor can inhibit the interaction between GBM and cells of the immune innate system including tumor-associated macrophages. Studies on murine models showed that macrophages develop resistance to CSF1 inhibition through the expression of the insulin growth factor 1 . The co-In the last years, several agents targeting angiogenesis have been tested in GBM without significant results. The only FDA-approved drug is bevacizumab which has been associated with prolonged progression-free survival, a reduction of symptoms related to the tumor but failed to show a significant impact on overall survival –136. SevSince GBM cells adopt extracellular vesicles to communicate with surrounding tissue, a system able to reduce vesicles uptake and synthesis can be a promising target. Pre-clinical studies showed that inhibitors of neutral sphingomyelinase (GW4869) can reduce the production of extracellular vesicles while heparin and annexin A1 inhibitors can reduce vesicles intake by target cells –153.Direct inhibition of junctions between GBM and microenvironment is also a strategy under investigation. Inhibition of connexin 43 is essential to inhibit the interaction between GBM and astrocytes . AnotherGBM remains a fatal disease with limited treatments. The reason for this failure could be partially explained by the development of a complex and effective net of interactions between tumor cells and surrounding tissue cells. The microenvironment resulting from the manipulation carried out by cancer cells can feed and stimulate GBM proliferation hiding and protecting tumor cells from systemic treatments.Interactions between tumor, endothelial and immune cells are under careful assessment due to the availability of drugs targeting these pathways. Recently, also the associations between glioblastoma and astrocytes, oligodendrocytes, white matter, and neurons come out from the shadow offering novel promising targets with therapeutic implications.Due to the presence of deep communications between tumor cells and the microenvironment the use of agents targeting more than one altered intracellular cascade at the same time appears a promising approach.In this optic novel immune-combination including immune checkpoint inhibitors) targeting PD-1/PD-L1, IDO1, TIM3, LAG3), engineered immune cells (CAR-T and CAR M), immune agonists , and BET inhibitors are under investigation. Notably, tumor-associated macrophages are one of the most important cell-associated with the development of an immune-depressive habitat. Thus, a strategy able to reverse this effect such as CAR M assumes a particular interest.Strategies aimed to inhibit the signaling between GBM cells and the microenvironment are also of key importance as able to inhibit the manipulation carried out by tumor cells on surrounding tissues. Inhibitors of sphingomyelinase, annexin A1 can act on extracellular vesicles secretion and intake while other agents such as ADAM10 inhibitors could directly interfere with the jap junctions connecting GBM to other cells.Finally, it should be noted that a specific microenvironment composition (such as that described on the white matter) can promote GBM differentiation and convert a tumor cell into an oligodendrocyte well-differentiated element. This surprising finding suggests that there are some elements inside the microenvironment that can provide inhibitory messages to GBM reversing its natural course. Interactions between neurons and glia  appear of particular interest and should be further assessed as could hide important targets for novel drugs development. In conclusion, GBM should be considered as a network of interactions in which an action perpetuates against tumor cells result in a response of the associated microenvironment and vice versa. The role of the microenvironment should be always considered during pre-clinical studies and would offer novel targets for patients with GBM.VN and LG: writing and draft. EF, AB, AT, and SB: project conception and reviewing. 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.


In transplantation, donor and recipients frequently differ in age. Senescent cells accumulate in donor organs with aging and have the potential to promote senescence in adjacent cells when transferred into recipient animals. Characteristically, senescent cells secrete a myriad of pro-inflammatory, soluble molecules as part of their distinct secretory phenotype that have been shown to drive senescence and age-related co-morbidities. Preliminary own data show that the transplantation of old organs limits the physical reserve of recipient animals. Here, we review how organ age may affect transplant recipients and discuss the potential of accelerated aging. Organ transplantation is the treatment of choice for end-stage-organ failure. The supply of organs, however, is limited, resulting in prolonged waiting times with many patients dying or becoming too ill to be transplanted. Aging demographics have incrementally affected the deceased donor population with older donors showing the by far largest proportional increase. Organs from older donors are, at the same time, underutilized, frequently discarded or not even considered .The most obvious strategy that may close the gap between demand and supply may thus be an optimized utilization of older organs from deceased donors , 3. IncrSenescent cells accumulate with aging and have been identified as critical in driving the immunogenicity of older organs linked to the accumulation of cell-free mitochondrial-DNA that accelerate alloimmune responses . Recent Here, we introduce potential mechanisms and consequences of prompting bystander senescence and discuss clinically relevant aspects of senescent cell spread when transplanting older organs. Although speculative, age-disparate transplantation may also provide unique opportunities as the transplantation of young organs may contribute to rejuvenation.Cellular senescence is characterized as a stable and terminal state of growth arrest based on acquired anti-apoptotic pathways  that render senescent cells resistant to apoptosis . Thus, sSenescent cells exhibit a distinct, pro-inflammatory secretome consisting of cytokines , chemokines  and matrix remodeling enzymes termed the “Senescent Associated Secretory Phenotype” (SASP) . The proINK4a facilitates cell cycle arrest and can therefore also be used as a senescent cell marker . Thus, continuous exposure of intact fibroblasts to senescent cells resulted in increased numbers of DNA double-strand breaks (DSBs) indicating senescence, which had been inhibited when blocking gap junction-mediated cell–cell contact  displays an inevitable feature of organ transplantation promoting a sterile inflammation linked to the release of various pro-inflammatory cytokines coinciding with the production of SASP by senescent cells. It appears thus possible that IRI may aid to the promotion of senescence in transplant recipients.via p53 activation and the induction of micro-RNAs, effects that further amplify mitochondrial dysfunction , BCL-2 inhibitors such as navitoclax that inhibit the pro-survival pathway BCL-2/BCL-xL, ii), PI3AK/AKT inhibitors including Dasitinib and Quercetin (D & Q) that constrain the synergistic effects of PI3K and Akt inactivating the apoptosis mediating proteins Bad, caspase-9 and FKHRL1 and iii), FOXO regulators such as Foxo4-DRI interfering with the inhibition of p53-mediated apoptosis . In addi+ Atpase  while mo+ Atpase  , for instance, containing RNAs, proteins, and lipid components that may be released by transplanted tissue stem cells, have been shown to promote stem cell plasticity and tissue regeneration –82. Notain vitro .At least in theory, adult stem cells that have been detected in multiple recipient tissues including muscle, liver, myocardium and endothelium following hematopoietic stem cell transplantation –86 may sThe transplantation of older donor organs is associated with more frequent acute rejection rates and compromised outcomes calling for age-specific treatment approaches that may improve the quality of older organs while reducing immunogenicity. At the same time transplanting an organ from an old donor may pose a risk for young recipients. The clinical relevance and significance of this concept appears obvious and demands therefore a thorough evaluation. The potential of rejuvenation when transplanting young donor organs, in turn, are of theoretical consideration and will also demand a detailed analysis. Critical will therefore be studies that probe either accelerated aging or rejuvenation in relevant pre-clinical transplant models utilizing immunosuppression.Optimizing the use of available organs for transplant including those from older donors continues to be of critical importance to close the gap between demand and supply in organ transplantation. Utilizing senolytic drugs that selectively deplete senescent cells may constitute a potential approach to improve the outcomes of older organs while restricting the spread of senescence.All authors listed have made a substantial, direct, and intellectual contribution to the work and approved it for publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Lymphatic system is identified the second vascular system after the blood circulation in mammalian species, however the research on lymphatic system has long been hampered by the lack of comprehensive imaging modality. Nanomaterials have shown the potential to enhance the quality of lymphatic imaging due to the unparalleled advantages such as the specific passive targeting and efficient co-delivery of cocktail to peripheral lymphatic system, ease molecular engineering for precise active targeting and prolonged retention in the lymphatic system of interest. Multimodal lymphatic imaging based on nanotechnology provides a complementary means to understand the kinetics of lymphoid tissues and quantify its function. In this review, we introduce the established approaches of lymphatic imaging used in clinic and summarize their strengths and weaknesses, and list the critical influence factors on lymphatic imaging. Meanwhile, the recent developments in the field of pre-clinical lymphatic imaging are discussed to shed new lights on the design of new imaging agents, the improvement of delivery methods and imaging-guided surgery strategies. Blood and lymphatic vessel networks are two central components of cardiovascular system in vertebrate, playing a pivotal role in body homeostasis maintenance and involving in the development of multiple diseases . CompareAs a significant transportation route for various soluble antigens and activated antigen-presenting cells from peripheral tissues to the draining lymph nodes, the immune cells in the lymphatic system undertake the important immunosurveillance functions . LymphatThe lymphatic system is characterized by slow unidirectional flow, colorless transparency and minor diameter, which make it difficult to be recognized even under microscopy . TherefoNanomaterials exhibit unparalleled advantages in the field of theranostic bioapplications benefiting from their excellent properties, including tunable structures, facile modifications, size effect and sensitive stimuli-responsiveness –15. The In this review, we summarize the established approaches of lymphatic imaging used in clinic and introduce their strengths and weaknesses Fig. . SeveralNanoparticles used for lymphatic imaging usually referred to the size of 5–100 nm, locating in the “just-right” region for lymphatic drainage. The subtle size endows nanoparticles with prolonged retention in circulatory system, enhancing imaging resolution and pharmaceutical effect, rather than being trapped in the capillaries of the organs. Actually, both the characteristics of nanomaterials and lymphatic structure have great influence on the imaging results. In this section, we mainly introduce the special characteristics of lymphatic structure and the strategies for the design and delivery of nanoparticles.The lymphatic capillaries are ducts with a closed blind end, which consist of single-layer LECs and incomplete basement membrane . Unlike It is widely recognized that the size of the nanomaterials determines its selective entrance in blood or lymphatic vessels. Typically, nanoparticles 5 to 100 nm in diameter are suitable for lymphatic drainage, depend on the maximum gap ca.120 nm) between the endothelial cells of lymphatic capillary , 56. How0 nm betw. demonstrated that the same size of NPs but different zeta potentials  showed different cellular uptake, the cationic NPs exhibited higher cellular uptake than the other two in eight different cell lines consisting of epithelial cells, fibroblastic cells, carcinoma and megakaryoblastic cells –8599mTc-. reported a 99mTc-labeled biomineralization nanoparticle for multimodal imaging of lymphatic drainage and network in real-time using SPECT/MRI and NIR fluorescence. The gadolinium oxide nanoparticles were synthesized by the biomineralization approach from Gd and ovalbumin, in which the radioactive tracer and cypate was conjugated to ovalbumin scaffold. The corresponding MRI/NIR/SPECT nanoparticles (99mTc-Gd@OVA-Cy) with the appropriate size exhibited high resolution and sensitivity lymphatic imaging .Fi.Fi. inno. developed a cell membrane-anchored ratiometric upconversion nanomaterial, UCNPs-Cy3/Pep-QSY7/Ab, to visualise the MMP secretion in situ, and further applied in imaging metastatic SLNs. Cy3 and MMP2 substrate peptide labelled by QSY7 quencher (Pep-QSY7) were decorated on the surface of UCNPs. The UCNPs emission at 540 nm was quenched by QSY7 due to Cy3-mediated luminescence resonance energy transfer (LRET) process. Epidermal growth factor antibody (Anti-EGFR) was conjugated to UCNPs via a PEG44 linker for specific anchoring on the membrane of tumor cells. When MMP2 was secreted by tumor cells, Cy3 emission at 580 nm could be recovered upon MMP2-responsive Pep-QSY7 cleavage. So, the MMP2 secretion monitoring system anchored on the cell membrane achieved visualization of the MMP in metastatic SLNs in vivo . A. A. deverecisely , 141. Asrecisely –145.Fig.. successfully modified an extant NIR-I molecule whose emission extend to NIR-II (1200 nm) by converting heteroatomic sulfur to selenium in the receptor unit of D-A-D-type AIEgens. Nanoparticles named as L897 NPs encapsulating the NIR-II emissive AIEgens realized non-invasive lymphatic system imaging with high signal-to-background ratio. The fluorescent emission tail of this nanomaterial extended to 1200 nm and the quantum yield ascends to 5.8%. When L897 NPs were intradermally injected, the popliteal and sacral lymph nodes were quickly lit up and the lymphatic vessels between them were also clearly visualized .Fi.Fi. succ. reported a multimodal and multicolor nanosized imaging agent, which coalesced 111In-labeled radionuclide and five-color optical dyes to visualize the local lymphatic drainage and detect SLNs. Radionuclides allowed absolute quantification and augmented penetration depth, while multicolor optical imaging described different drainage regions with superior spatiotemporal resolution [. developed NaLuF4-based multifunctional upconversion nanoparticles doping with Gd3+, Yb3+, Er3+ and Tm3+. Lutetium (Lu) showed strong X-ray absorption for CT imaging, lanthanide performed excellent upconversion luminescence signal with unique properties, such as sharp emission lines, large stokes shifts, superior photostability, and non-blinking. Gd3+ provided outstanding T1-weighted MRI signal and relaxation. Studies had shown that the relaxivity parameter (r1) could reach as low as 2.273 s−1 mM−1, which was half of commercial T1-enhanced MRI agents while the signals acquired was higher than most of the reported Gd-doped nanoparticles , inorganic nanoprobe integrating PET and fluorescence dual-mode imaging modalities . Bradbur68 Ga-NEB to evaluate the feasibility of combining PET/MRL with FL (ICG) in rapidly visualizing lymphatic system and preoperative staging in lymphedema. The results proved that 68 Ga-NEB could provide detailed three-dimensional structures of central and peripheral lymphatic system with higher temporal resolution. Moreover, the picture of impaired lymphatic vessels was clearly revealed by combining PET/MRL and fluorescence lymphangiography. These multimodal images had important implications for evaluating lymphedema severity and staging damaged lymph vessels, thus contributing to make an individualized treatment plan , which can self-assemble in vivo with desirable biological functions  and antigens traffic from tissues to draining lymph nodes, where the lymphocytes execute the function of immune monitoring. From this perspective, more functional information about the lymphatic system will be found by using multimodal imaging strategies to track immune cells in the lymph at different levels. Chen et alons Fig. . There wIn this review, a series of fantastic research about the design strategy of lymphatic nanoformulations and lymphatic imaging modalities have been summarized. The individual modalities based on various creative nanomaterials discussed above had express high-quality information of lymphatic drainage. As an ideal lymphatic contrast agent, the following characteristics should be taken into consideration:To better access the function of lymphatic, it is imperative to integrate multiple advanced lymphatic imaging modalities. Multimodal imaging technology with PET as the fundamental pillar has been widely developed in clinical practice in addition to lymphatic imaging, which may be related to lacking of fewer available clinical lymphatic contrast agents and the specific anatomy of peripheral lymphatic vessels. Represented by near-infrared fluorescence and photoacoustic imaging have brought light to the mentioned predicament above. It is envisioned that PET-CT (MRI)/optical multimodal imaging is promising become the best combination in clinical diagnosis and treatment in the future.There are a lot of important factors need to be considered in precise imaging of LNs including the locations of LNs , the characteristics of LNs (immunity or tumor metastatic), the injection practise of contrast agents (intravenous or subcutaneous injection) and so on. In order to specifically deliver imaging agents to targeting LNs, a major consideration emphasizes the difference of individual LNs microenvironment. In particularly, there are several options available for targeting the corresponding LNs such as the specific antigenic peptides presented on the surface of target cells, the lower pH or higher level of ROS/GSH in tumor/inflammation extracellular microenvironment, the aggregation of tumor-associated immune cells, excessive cytokine or enzyme release associated with tumor/inflammation etc. These factors described above can be used as stimuli during the preparation of nanoformulations for imaging LNs precisely.Surgery remains the mainstay of therapy for multiple primary diseases. Clinicians in the worldwide are now faced with the dilemma: how to avoid the iatrogenic injury and meanwhile make sure radical treatment? Imaging-guided accurate intraoperation navigation provides a feasible solution to address the problem. Given that the lymphatic system is characterized by colorless transparency and minor diameter, the lymphatic imaging with high spatiotemporal resolution could present accurate information of lymph nodes and lymphatic vessels in tumor radical resection and lymphatico-venous anastomosis, maximally shortening the operation time, improving the process efficiency and avoiding iatrogenic injury. Certainly, synthesizing nanomaterials with excellent properties and choosing a suitable multimodal imaging method are the first and key step to achieve these goals. Over the past decades, the research on lymphatic system is far less advanced than other systems owing to lacking of powerful means to visualize it comprehensively. As the interesting of lymphatic imaging continues evolve, it is imperative to look for innovative nanomaterials to overcome the obstacle by taking full advantage of individual element with distinct characteristics and eventually achieve clinical translation. We anticipate that an overall impressive description of the central-peripheral-superficial lymphatic system can be depicted at one time in the not-too-distant future and the lymphatic system is bound to return to the spotlight.