The Enhanced Recovery After Surgery (ERAS) or Fast Track is defined as a multi-disciplinary, peri- and post-operative approach finalized to reduce surgical stress and simplify post-operative recovery. It has been introduced more than 20 years ago by Khelet to improve outcomes in general surgery. Fast Track is adapted to the patient’s condition and improves traditional rehabilitation methods using evidence-based practices. Fast Track programs have been introduced into total hip arthroplasty (THA) surgery, with a reduction in post-operative length of stay, shorter convalescence, and rapid functional recovery without increased morbidity and mortality. We have divided Fast Track into three cores: pre-, intra-, and post-operative. For the first, we analyzed the standards of patient selection, for the second the anesthesiologic and intraoperative protocols, for the third the possible complications and the appropriate postoperative management. This narrative review aims to present the current status of THA Fast Track surgery research, implementation, and perspectives for further improvements. By implementing the ERAS protocol in the THA setting, an increase in patient satisfaction can be obtained while retaining safety and improving clinical outcomes. Enhanced Recovery After Surgery (ERAS) or Fast Track, is defined as the multi-disciplinary, peri- and post-operative approach performed to optimize patient’s health condition, reduce surgical stress, and improve post-operative recovery. It was introduced more than 20 years ago by Khelet et al.  to improERAS protocols have been developed to promote a shorter convalescence and a faster functional recovery without an increase in the rates of morbidity and mortality; it implies the promotion of a reduced post-operative hospital length of stay (LOS) ,4. An apTotal hip arthroplasty (THA) places a heavy financial load on the healthcare system, and it is, therefore, essential to optimize the available resources to speed up recovery and cut down the length of stay (LOS) without sacrificing outcomes . Fast TrERAS protocols should be based on the principle of “first better, then faster” . The aimThe last review on the subject was done by Wainwright et al.  over 3 yThe first key step in THA surgery performed according to Fast Track protocols is patient selection. Traditionally, patients referred to a Fast Track protocol were young and fit. However, these limitations have been progressively overcome because of the implementation of minimally invasive surgical techniques and optimized anesthesiologic procedures; therefore, more patients are nowadays considered suitable for ERAS protocols in THA . UltimatPatient motivation and awareness are key to the implementation of ERAS protocols: not all patients may feel safe in being discharged early from the hospital. The psychological health of the patient influences physical recovery following THA surgery ; therefoAn additional key element that must be discussed with the patient in preoperative counseling is the presence of discharge criteria. These allow standardization of treatment goals and ensure that patients are discharged only when in a safe condition. Discharge criteria should be the following: stable ambulatory autonomy with crutch support and without dizziness; the absence of nausea or vomiting or a minimal presence that can be controlled without pharmacological intervention; pain <3  at rest and <5 during ambulation; and bleeding compatible with a normal postoperative course that does not require repeated dressing changes ,38.Smoking is a known risk factor associated with a high rate of perioperative and postoperative complications, like myocardial infarction, cardiac arrest, pneumonia, urinary tract infection, sepsis, acute renal failure, and mortality ,38,39. TLike smoking, alcohol use can negatively influence patient outcomes after surgery . When abPre-operative anemia in the orthopedic setting is extremely common, with a prevalence ranging from 15 to 40% . Anemia Reduced LOS requires rapid rehabilitation to allow the patient to regain functional autonomy. Traditionally, physical therapy is performed in the postoperative setting, and the patient regains the ability to walk and climb stairs with the assistance of a physical therapist that teaches the basis of walking with crutches, postoperative exercises, and avoidance of movements promoting dislocation. Several studies have recently shown that learning how to use crutches before THA performance and preparing the body with appropriate exercises before surgery decreases the time to independence, reduces LOS, and improves patients’ confidence ,58. ThesMalnutrition is a risk factor for postoperative complications; it slows tissue repair, extends in-hospital LOS, determines longer recovery times, and is associated with increased mortality . IndicatDiabetes is a common comorbidity among obese patients. A pre-operative work-up is of the utmost importance in trying to reduce HbA1c below the 8% threshold associated with a higher incidence of wound complications and infections .Optimization of the anesthetic protocol is a major issue in patient candidates for Fast Track THA surgery. It requires specific team skills and a careful preoperative workup to support the patients in all the phases of the in-hospital stay. General and spinal anesthesia are the most common form of anesthetic protocols performed in this setting . DiffereTo improve postoperative pain after THA, multimodal-analgesia can be implemented and opioid therapy dosage can be reduced by performing peripheral regional nerve blocks (PNB) . PNB is Two other techniques that can take place in pain control are suprainguinal fascia iliaca block and pericapsular nerve group block (PENG), with the latter resulting in better motor functions ,82, perfPeri-capsular infiltration is still a debated topic as a recent RCT showed no effect of local infiltration analgesia after THA . More evTHA can be performed by different surgical approaches. Despite the recent trend toward minimally invasive muscle-sparing approaches , there iAntifibrinolytic agents have gained popularity to reduce bleeding and allogenic blood transfusions and, among them, tranexamic acid (TXA) certainly is the most extensively used ,95. TXA Multimodal analgesia for orthopedic surgery is now widely practiced as a means to reduce opioid use and opioid-related side effects. A multimodal approach is likely to produce superior analgesia than an opioid-only-based approach because multimodal analgesic agents target a variety of pain pathways . AlthougThe use of a perioperative systemic glucocorticoid, although having robust evidence in knee replacement, is still debated in elective hip arthroplasty . Their eOne of the problems associated with THA is the high incidence of an immediate postoperative orthostatic intolerance, which is defined as a drop >20 mmHg in systolic blood pressure or >10 mmHg in diastolic blood pressure associated with symptoms including dizziness, nausea, vomiting, vision disturbances, and syncope. This biological reaction, driven by an unknown impaired vasomotor response, has an incidence of 40% at 6 h and 20% at 24 h postoperatively , and it It is well known how these three complications slow the postoperative rehabilitation course; these are mostly attributable to the use of opioid drugs, and support the discontinuation or avoidance of opioid drugs in Fast Track multimodal analgesic protocols . Of the Postoperative urinary retention (POUR) is a common complication that occurs in more than 40 percent of patients subject to THA, especially when spinal anesthesia is performed. Although multiple pharmacological approaches are commonly used to prevent this event, when POUR occurs catheterization is required, either indwelling or intermittent ,122. ThoInternational guidelines on antithromboembolic prophylaxis are clear in suggesting extensive 10–30-day prophylaxis in high-risk procedures, including major orthopedic surgeries ,132,133.Early function and mobilization are key factors for the success of a short-stay program, and recent Fast Track guidelines recommend that patients should be mobilized as soon as possible after surgery . MobilizTo maximize patient satisfaction and surgical outcomes while reducing costs and optimizing resources, Fast Track protocols were progressively introduced in THA surgery: a multi-disciplinary approach in the peri- and post-operative period, tailored to every patient .Several studies have shown the advantages of the implementation of Fast Track protocols. Maempel et al.  studied Patients operated on according to ERAS protocols with early discharge from the hospital have long been considered at risk of subsequent re-admission, typically for surgical site infection, dislocation, deep vein thrombosis, or symptomatic anemia . HoweverPatient selection was the critical issue in ERAS protocols, being more selective in the early years , but gotFast Track protocols are not without limitations. Patients need to be involved and well aware of the procedure. For this reason, patients with psychiatric conditions that may impede cognitive function and elderly patients with dementia are not eligible. Moreover, people with serious coagulation disorders or systemic diseases that may call for intensive care, numerous transfusions, or dialysis must be removed from the program. Another reason for exclusion could be the lack of nearby medical services or transportation issues since social and logistical support is of paramount importance in the period close to surgery to ensure a consonant rehabilitation pathway .Fast Track practices can be implemented by an innovation introduced to cope with the COVID-19 pandemic. By reducing LOS, the use of Fast Track practices invariably results in reduced health professional-patient contact, just as during the outbreak. A more detached relationship is not associated with a reduction in clinical outcomes however, patients report a reduction in overall satisfaction . An implIn conclusion, Fast Track THA surgery is effective in improving patient outcomes and satisfaction while decreasing healthcare costs and utilization of available resources. Optimization of the patient’s health preoperatively is crucial, together with appropriate patient selection in terms of general health status, motivation, commitment, and logistics. Pre-habilitation and customized post-operative rehabilitation protocols are also priorities. Future research will outline even more aspects to fine-tune ERAS protocols in THA candidate patients to minimize complications and costs while maximizing patient outcomes and satisfaction.


Wound healing is a long-term, multi-stage biological process that mainly includes haemostatic, inflammatory, proliferative and tissue remodelling phases. Controlling infection and inflammation and promoting tissue regeneration can contribute well to wound healing. Smart biomaterials offer significant advantages in wound healing because of their ability to control wound healing in time and space. Understanding how biomaterials are designed for different stages of wound healing will facilitate future personalized material tailoring for different wounds, making them beneficial for wound therapy. This review summarizes the design approaches of biomaterials in the field of anti-inflammatory, antimicrobial and tissue regeneration, highlights the advanced precise control achieved by biomaterials in different stages of wound healing and outlines the clinical and practical applications of biomaterials in wound healing. This article reviews the design methods of biomedical materials in the field of anti-inflammatory, anti-bacterial and tissue regeneration.Biomedical materials have precise control in different stages of wound healing.Biomedical materials provide new insights into the treatment of cutaneous injury and the current challenges.Normal wound healing is a complex physiological process with haemostatic, inflammatory, proliferative and remodelling phases. After wound formation, the coagulation system is activated and a local fibrin clot forms . This isBurns, diabetes, infections and inflammation may cause a wound to heal slowly or become chronic nonhealing. Owing to the complexity of healing, re-establishing post-wound skin function remains difficult. Dressings, devices, medications, surgical interventions and biological approaches are currently used to treat wounds. Although the therapeutic efficacy of these approaches has been demonstrated, they are expensive and can result in adverse reactions such as rejection, infection and allergy. The main disadvantage is that these methods have a limited ability to initiate healing without stimulating subsequent reconstructive phases.Therefore, innovative therapies should target specific events during the healing process to accelerate closure and restore healing tissues. Recently, Ag-modified polydopamine nanoparticles were embedded in a polysaccharide hydrogel-based network, resulting in excellent antimicrobial properties that can effectively promote wound healing . FurtherIn addition to nanomaterials, biomaterial-based wound dressings with biodegradability and biocompatibility are widely used at various stages of the wound-healing process . Biomatein situ tissue growth, promote tissue repair and respond to the internal environment [Biomaterials have been used in medicine since the 1950s . Most maironment .Controlling infection and inflammation, as well as promoting tissue regeneration, are key strategies for managing favourable wound healing. Most chronic wounds are associated with infection and microbial film formation, which can lead to amputation, sepsis or even patient death if left untreated . The immIn most cases, specific components, such as nanomaterials, growth factors, cytokines and hydrogels, can stimulate each stage of the wound-healing process. Several review articles have discussed recent advances and current clinical strategies . AlthougBiomedical materials can be classified into three categories based on their properties: metals, non-metallic inorganic materials and polymers. Metallic biomaterials are widely used in clinical applications owing to their excellent mechanical and regenerative properties ,29. TheyNon-metallic inorganic materials are classified into two main categories: crystalline and amorphous. Non-metallic and metallic elements are bonded together via covalent or ionic bonds to form bioceramic materials with immunomodulatory effects. For example, bioactive ceramics can regulate and convert M1-type macrophages into M2-type macrophages during the inflammatory phase of wound repair by creating a favourable microenvironment .Bioactive glasses are another important class of non-metallic inorganic biomaterials that cause biological effects by binding to bone and releasing bioactive ions such as silicon, calcium and phosphate. Recently, biomaterials based on bioactive glasses have been optimized for various biological functions ,44.There are numerous polymer types with various properties, including hydrogels, nanomaterials, bioderived materials, natural polymers, synthetic polymers and biopolymers ,46. HydrNumerous multifunctional nanomaterials have been used in wound therapy in recent years to promote wound healing by taking advantage of their high reactivity, specific surface area, catalytic efficiency and loading efficiency. For example, iron tetroxide nanoenzymes have reaction kinetics similar to those of natural enzymes and can convert hydrogen peroxide into highly toxic hydroxyl radicals under mild conditions, resulting in antibacterial effects. Nanoenzymes have the advantages of low cost, easy synthesis, high stability and no external energy input compared with natural enzymes, and have become effective antimicrobial agents for wound treatment and bacterial infections .Biologically derived materials mainly include the extracellular matrix and vesicles. Previous studies have demonstrated that mesenchymal stem cell (MSC)-derived extracellular vesicles have favourable repair effects on burns and diabetic wounds ,54. NatuLacerations, burns and chronic refractory wounds are common clinical complications. Any disturbance in the wound-healing process, such as internal or external environmental irritation that results in chronic wound or prolonged healing, can pose a significant burden to clinical care and patients. Therefore, biomaterial design should be based on the principles of reducing healing time and improving healing efficiency. Anti-inflammatory, antibacterial and proliferation-promoting activities are three aspects of biomaterial design that need to be considered , and a dIn recent decades, tremendous progress has been made in the research of biomaterials based on wound healing, which has contributed greatly to the application of biomaterials in the field of tissue engineering . CurrentVarious antimicrobial dressings have been extensively studied to promote wound healing. However, comparing dressings is difficult because dressings with different structures cannot simultaneously meet the requirements of different wounds, which requires researchers to study the wound-healing effects of dressings more specifically. In addition, the inevitable use of organic solvents and cross-linking agents in the preparation of dressings with specific morphologies necessitates a critical evaluation of material biocompatibility before clinical application. Finally, the development of high-volume production facilities and timely and effective antimicrobial biomedical materials is important for promoting wound healing. This necessitates the use of cost-effective materials and relatively simple synthetic processes to manufacture dressings.Inflammation is a tightly regulated process that aims to eliminate pathogenic damage by removing damaged tissues and restoring tissue homeostasis ,69. UncoBased on their physical structure and ability to bind various bioactive molecules, biomaterials have been developed and used to modulate macrophage function during the inflammatory phase of wound healing, thereby locally manipulating the inflammatory response . In gene3O4) nanoparticles loaded with fibroblast growth factor (bFGF) using a simple mussel-inspired surface immobilization method that stabilizes bFGF under various conditions and exhibits sustained release of bFGF. In vivo and ex vivo studies have demonstrated that Fe3O4 nanoparticles loaded with bFGF can significantly promote wound healing by polarizing M2 macrophages and promoting cell proliferation  nan+ and serine proteins to form a dual-network hydrogel that could modulate macrophage polarization and promote wound repair in diabetes [Glycyrrhizic acid (GA) is a natural compound derived from the roots of liquorice plants that has the potential to be an ideal building block for advanced hydrogel dressings with intrinsic immunomodulatory activity. GA and its derivatives have previously been used to treat chronic inflammatory conditions. In one study, hydrogels were loaded with GA that was cross-linked with Zndiabetes .In addition, temperature-sensitive chitosan (CS) hydrogels have been used to load prostaglandin E2 (PGE2) bio-signals with pro-inflammatory immunomodulatory effects . The CS There are still challenges to overcome in the field of tissue repair and regeneration, including complex material fabrication, uncontrolled drug release, high costs and drug-related side effects; however, various drugs, cytokines and cell delivery in hydrogels are helping to advance the field . RecentlIt has also been shown that when the surface groove of the material is 1–5 μm, it is difficult to induce an inflammatory response in the body to regulate the conversion of macrophages to the M2-type, whereas when the surface of the material is sharp, it is easy to induce an inflammatory response in the body . Thus, btert-butyl alcohol, lyophilized them to obtain an aerogel, and heat-treated them to stabilize the structure, after which a hydrophobic layer was electrospun on top. The aerogel accelerates diabetic wound healing by synergistically shortening the inflammation period  and release them into the inflamed tissue. Microenvironmental ROS selectively leads to the activation of transcription factors that control the expression of pro-inflammatory cytokines ), chemokines and protein hydrolases, including Matrix Metalloproteinases (MMPs) and serine proteases.During the inflammatory phase of the wound, local tissue blood vessels are constricted and the blood supply is insufficient, resulting in a lack of nutrition and oxygen and an increase in glycolysis, which raises lactic acid and PaCO2-FasL) was designed  light illumination owing to the photothermal effect of polydopamine nanoparticles, and was able to not only synergistically sterilize but also achieve precise release of antibiotics after NIR light illumination [Antibiotics have been used to treat pathogenic bacterial strains since the 1940s ,108. Antmination .et al. prepared a novel antimicrobial hydrogel wound dressing [in vivo experimental results demonstrated that the hydrogel effectively promoted the wound-healing process.The broad-spectrum antimicrobial activities of cationic polymers and nanoparticles make them popular among researchers. Pathogens can be effectively inactivated by electrostatic interactions between positively charged polymers and negatively charged bacterial cell membranes. Cationic polymers contain networks of positively charged groups, can be composed of natural or synthetic organic polymers and have been used to construct antimicrobial biomaterials . For exadressing . The hydSeveral nanoparticles have been studied for potential antimicrobial applications, including those containing silver, copper, gold and other metals ,114. HowS. aureus [Recently, antimicrobial peptides (AMPs), which are non-toxic and non-resistant small-molecule peptides, have been proposed for treating drug-resistant bacterial strains . Researc. aureus . HoweverIn addition to a material carrying antimicrobial agents to achieve bactericidal effects, the material itself can be modified to induce bacterial cell membrane rupture by allowing the pathogen to come into direct contact. For example, graphene oxide nanosheets can use the ‘nanoblade effect’ to break the lipid bilayer of bacteria .S. aureus biofilms without using biocides, resulting in a 50–80% reduction in colonization rates [In addition to their unique morphology, the wings of cicadas and dragonflies exhibit antimicrobial activity because of the nanoscale columnar structures on their surfaces. Inspired by their unique structures, several synthetic bio-nanomorphic surfaces with similar nanomorphologies have been used for effective bacterial killing . Other ron rates . The antS. aureus adhesion on the stainless steel surface, but the nanoweb structure was superior to the nanostrip array structure in terms of antibacterial adhesion performance. The hydrophobic properties of this material can also be used to create antibacterial agents [Femtosecond lasers have also been used to produce nanostrip arrays and nanoweb structures on the surface of stainless steel . Both nal agents .Furthermore, superhydrophobic surfaces can inhibit bacterial adhesion owing to the mutual repulsion between the surface and the hydrophobic surface of bacteria. Bacterial contact and adhesion are reduced on superhydrophobic surfaces because of the adsorption of an air film on the surface, which reduces the solid area fraction, resulting in a self-cleaning ability of the surface, which can effectively remove initially-adhered bacteria in subsequent cleaning. However, these changes in the physical signals of the material have limited bactericidal effects.S. aureus, E. coli and Candida albicans under NIR light  [The recent development of photothermal therapy (PPT) and photodynamic therapy (PDT) has resulted in significant advances in biomaterials for antimicrobial purposes. PPT is a safe and effective strategy for combating infections using appropriate photothermal materials that can convert light into heat and kill bacteria via cell wall degradation and protein/enzyme denaturation. For example, in one study, a polymer substrate immersed in a weakly alkaline dopamine solution resulted in a polydopamine photothermal antimicrobial coating on its surface that exhibited excellent photothermal antibacterial properties against  W·cm−2) . PPT is  W·cm−2) ,127.1O2) and hydroxyl radicals. The generated ROS can damage pathogen cell membranes and DNA [PDT is another promising strategy for precisely treating persistent bacteria remotely. PDT requires three factors: photosensitizers (PS), light sources and oxygen. When PS are irradiated with light at a wavelength matching the maximum absorption, the excited-state PS can generate toxic ROS, including superoxide, singlet oxygen  that may modulate wound healing at all stages and provide a balanced environment throughout the wound healing process, thereby reducing potential complications. Many of the therapeutic effects of biomaterials utilize a complex of materials with different functions that act on different stages of wound healing, thus achieving a synergistic promotion of the process.In addition to macrophages, T cells play an important role in regulating the inflammatory response during the inflammatory phase of trauma repair. γδ T cells in the skin have been shown to provide the necessary cytokines and growth factors that regulate intraepidermal homeostasis, but also promote inflammation at the wound site. Dermal γδ T cells provide cytokines and chemokines that regulate inflammation throughout the skin and feed back to epidermal γδ T cells ,161. GivThere are many requirements for the design of materials to promote wound healing, such as the need for good adhesion and haemostatic properties. Recently people are also investigating the wound detection functions of hydrogels, such as can be achieved by adding some conductive biomaterials to the hydrogel . Such biThis paper reviews the design approaches of biomedical materials for anti-inflammatory, antibacterial and regeneration promotion. Anti-inflammation is an important strategy in the inflammatory phase of wound healing, and the design of anti-inflammatory biomaterials must include the slow release of relevant molecules, changing the physicochemical properties of the biomaterials or using biomaterials to mediate relevant signals in the trauma microenvironment. This in turn leads to the conversion of macrophages in the trauma into M2 type macrophages, and future research should also consider using other immune cells to achieve better control of inflammation. Antimicrobials are also an important tool in improving healing efficiency. The key to the design of common antimicrobial materials is to use the material as a carrier to carry/immobilize the antimicrobial signal; bactericidal effects can also be achieved using the material’s own properties through physical/chemical modifications of the surface of the material. The synthesis of more efficient and economical antimicrobial materials is a problem that needs to be addressed in the future. The key to accelerating the proliferation and remodelling phase of wound healing is to use the carrier slow-release properties of biomaterials to provide the relevant proliferation and regenerative active molecules to synergistically promote wound repair. The choice of relevant active molecules largely affects the efficiency of wound healing and so exosomal synergistic biomaterial therapy has become a new therapeutic trend.


Individuals diagnosed with neurodevelopmental conditions such as autism spectrum disorder  often experience tissue inflammation as well as gastrointestinal dysfunction, yet their underlying causes remain poorly characterised. Notably, the largest components of the body’s immune system, including gut-associated lymphoid tissue (GALT), lie within the gastrointestinal tract. A major constituent of GALT in humans comprises secretory lymphoid aggregates known as Peyer’s patches that sense and combat constant exposure to pathogens and infectious agents. Essential to the functions of Peyer’s patches is its communication with the enteric nervous system (ENS), an intrinsic neural network that regulates gastrointestinal function. Crosstalk between these tissues contribute to the microbiota-gut-brain axis that altogether influences mood and behaviour. Increasing evidence further points to a critical role for this signalling axis in neurodevelopmental homeostasis and disease. Notably, while the neuroimmunomodulatory functions for Peyer’s patches are increasingly better understood, functions for tissues of analogous function, such as caecal patches, remain less well characterised. Here, we compare the structure, function and development of Peyer’s patches, as well as caecal and appendix patches in humans and model organisms including mice to highlight the roles for these essential tissues in health and disease. We propose that perturbations to GALT function may underlie inflammatory disorders and gastrointestinal dysfunction in neurodevelopmental conditions such as autism. The gut-brain axis is the well-established, bi-directional communication between the gut and brain. Classically, the gut-brain axis influences a variety of communication pathways such as the neuroendocrine system, immune system and the peripheral nervous system ). However, recent evidence has shifted the classical concept of the gut-brain axis to include the microbiota, with many studies highlighting the microbiota playing a mediatory role in these pathways. The microbiota-gut-brain axis modulates the body’s response to changes in the microbiota within the gastrointestinal (GI) tract, influencing gut function, bodily immune response, physiological stress and cognitive behaviour. Several studies indicate these complex interacting pathways are disturbed in neurodevelopmental disorders including autism spectrum disorder  , which is altered in a mouse model of autism  and microscopic (such as the cellular composition and morphology of patches) are evident. Microbial populations are specific to gut regions and their interactions with surrounding environments likely influence the development of lymphoid aggregates in the small intestine and caecum of the host. As such, alterations in host, including changes in nervous system function and patterns of innervation, can influence the development and function of Peyer’s patches and caecal patch tissue. Furthermore, Liu and colleagues reported that host genetics could modify the microbiome  . Throughout foetal development in mice, chemokines such as CXCL12 and -13, CCL19, -20 and -21, and adhesion molecules including vascular cell adhesion molecule 1 (VCAM-1), mucosal vascular addressing cell adhesion molecule 1 (MAD-CAM1) and intercellular adhesion molecule 1 (ICAM-1), altogether signal to promote the circulation of LTi cells and the migration of T- and B-lymphocytes into primordia resembling Peyer’s patches  of the appendix  and T and B cells  that influences signalling primarily within the small intestine, whereas caecal patches produce IgA-secreting B cells that migrate both to the small intestine and the colon  cells are located in the follicular associated epithelium of Peyer’s and caecal patches and transcytose luminal content. M cells can be morphologically distinguished from intestinal enterocytes based on their short irregular microvilli located at the apical surface, in contrast to the uniform and densely packed microvilli of enterocytes.The intestinal epithelial cells in the follicular associated epithelium originate from stem cells in crypts positioned between a villus and dome of the Peyer’s patches Heath . Cells o-epithelial dome, antigen presenting cells interact with B and T lymphocytes, dendritic cells and macrophages to form part of the mononuclear phagocyte system. In this process, antigen presenting cells present the antigen/class II major histocompatibility complex (MHC-II) and cytokines to naïve lymphocytes. Within Peyer’s patches, B cells undergo an immunoglobulin class switch  due to the influence of factors such as transforming growth factor-β (TGF-β), and IL-10  have not been reported, microbial dysbiosis in faecal samples were identified  and that these cells had a more rounded morphology, potentially indicating an activated cellular phenotype (Sharna et al. Gastrointestinal dysfunction is a common co-morbidity observed in a range of neurodevelopmental disorders including ASD. GI symptoms commonly experienced by individuals diagnosed with ASD include constipation, diarrhoea, abdominal pain, and more (Holingue et al. GALT is an important component in sensing and responding to the microbial environment and therefore should be considered as a major component contributing to microbiota-gut-brain axis function. Alterations in the nervous system occurring in neurodevelopmental disorders may also impact GALT activity as well as gastrointestinal mucus production and composition, epithelial replenishment and the distribution of microbial populations along the gastrointestinal tract (Herath et al.


Patient-derived xenograft (PDX) models, which can retain the characteristics of original tumors in an in vivo-mimicking environment, have been developed to identify better treatment options. However, although original tumors and xenograft tissues mostly share oncogenic mutations and global gene expression patterns, their detailed mutation profiles occasionally do not overlap, indicating that selection occurs in the xenograft environment. To understand this mutational alteration in xenografts, we established 13 PDX models derived from 11 brain tumor patients and confirmed their histopathological similarity. Surprisingly, only a limited number of somatic mutations were shared between the original tumor and xenograft tissue. By analyzing deleteriously mutated genes in tumors and xenografts, we found that previously reported brain tumor-related genes were enriched in PDX samples, demonstrating that xenografts are a valuable platform for studying brain tumors. Furthermore, mutated genes involved in cilium movement, microtubule depolymerization, and histone methylation were enriched in PDX samples compared with the original tumors. Even with the limitations of the heterogeneity of clinical lesions with a heterotropic model, our study demonstrates that PDX models can provide more information in genetic analysis using samples with high heterogeneity, such as brain tumors. A brain tumor is one of the hardest tumors to cure, with a 5-year overall survival rate of less than 35%, even though many personalized therapeutic options have been developed . In 2016Large-scale genomic analyses, including The Cancer Genome Atlas project, provide information about molecular markers to group glioblastoma multiforme (GBM) into at least four molecular subtypes depending on genetic, epigenetic, and transcriptional alterations ,5,6. MorPatient-derived xenograft (PDX) models are considered a promising platform for personalized cancer research because they can recapitulate the genomic and epigenetic characteristics and microenvironment interactions during the progression of a brain tumor. The Mayo Clinic Brain Tumor PDX National Resource team analyzed the genetic alterations in 83 PDX samples from 24 patient tumors using whole-exome sequencing . They reA previous study reported discordance of the detailed mutation profile between xenograft and patient tumors with some sample dependency, ranging from 12% to 64% of shared mutations based on whole-genome sequencing . AnotherA recent study analyzing 15 trios of tumor-BTIC (brain tumor-initiating cell)-xenograft samples showed that some mutations were shared among the trio samples, but others were only identified in the xenograft model . Th. Th21]. We additionally investigated genes with somatic mutations enriched in the PDX samples or the original tumors. First, we examined 723 COSMIC cancer census genes commonly associated with various tumors a,b and iWe defined 409 genes with high-impact mutations  and investigated whether these gene mutations were enriched in tumors or PDX samples by subtracting the number of tumor samples with the mutated genes from the number of PDX samples with the same mutated genes e. These If brain tumor-specific mutations are enriched in PDX samples, we speculated that tumor-associated gene mutations that are not detected in the original tumor would be enriched in the PDX sample. To analyze enriched mutated genes in the tumor and PDX samples, we listed the individual enriched gene sets in each sample type. Genes with deleterious mutations were more diverse among the tumor samples, probably due to the heterogeneity of these samples.While 761 genes were selected as tumor-enriched genes, GO term analysis of the enriched mutated gene set of original tumors found no significant terms, probably due to low sensitivity owing to tumor heterogeneity. By contrast, we identified 1610 genes enriched in the PDX samples, and GO terms related to histone methylation, cilium movement, and microtubule depolymerization were significantly overrepresented . These gPDX models have been widely used to investigate tumors because they can mimic the in vivo environment where tumors grow. The rapid proliferation of tumor cells can help enrich tumors in this environment, which can help identify tumor-related mutations and to study their characteristics. However, although key driver somatic mutations were claimed to be well conserved between the original tumors and the PDX samples, comprehensive somatic mutations across genomes were not thoroughly investigated.Here, we analyzed the somatic mutation profiles of 13 PDX models from ten brain tumor patients and their original tumors. Even with this limited number of cases, the discordance of somatic mutations between the original tumors and the derived PDX samples showed that established PDX models may not recapitulate the genetic characteristics of the original tumors. Alternatively, insufficient genetic information might have been obtained owing to the high heterogeneity of brain tumors. Our sequencing depth was much lower (30×) than the recommended depth of whole-exome sequencing for the somatic mutation analysis of tumors (120×); therefore, it might be challenging to identify all somatic mutations in both sample types. A recent study suggests that patient-derived explants and glioma sphere lines derived from glioblastoma retain genetic similarity . HoweverThe cohort in our study is composed of small numbers and diverse pathologies of various brain-associated tumors , which show different biological behavior, may be the limitation of this study. We tried to establish more samples for this study, but the success rate was only 22% . AlthougWhen we analyzed the somatic mutations of genes related to brain tumor development, such as ARTX, BRAF, and MGMT, we identified more mutations in the PDX samples than in the original tumors a. FurtheWe further examined the somatic mutations of TP53  and EGFRPrevious studies showed the concordance of genome-wide functional features, such as copy number variations and gene expression patterns, between PDX samples and the original tumors. We also confirmed that the mutational signatures of each PDX sample were well matched with their original tumors. Additionally, two samples (SNPDX-037 and SNPDX-038) retained the mutational signatures caused by TMZ treatment. The patients who provided SNPDX-028 and SNPDX-045 had also been treated with TMZ, according to our records , but we Gene mutations related to histone methylation and ciliary functions were enriched among the PDX samples. A recent study showed that glioma stem cells (GSCs) derived from a GBM patient exhibited the suppression of ciliogenesis, which led to self-renewal rather than differentiation . When thAlthough our study demonstrated that the subpopulation of tumor cells might increase in our PDX models because of in vivo growth-promoting properties, further studies are required to investigate whether the mutations enriched in PDXs are important in the original brain tumors due to their associations with tumor-initiating cells or tumor stem cells. Alternatively, these mutations may increase cellular fitness in the xenograft environment, independently of their roles in tumor progression. Further analysis of more PDX models and paired primary tumor cell models, as well as single-cell level analysis, will help elucidate how to utilize and interpret the genetic features of PDX tumor models.


Vibrio cholerae, and this proportion probably varies by locationand over time [The Global Taskforce for Cholera Control (GTFCC) End Cholera Roadmap encouragescountries with endemic and epidemic cholera transmission to identify cholera hotspots inorder to prioritize multi-sectoral prevention and control strategies, including oralcholera vaccines and water/sanitation infrastructure . Althougver time . In otheV. cholerae O1 .Current World Health Organization (WHO) recommendations suggest that laboratoryconfirmation should be performed to confirm the start and end of outbreaks, and that‘it may be interesting to take a few samples randomly during the outbreak to makesure that the antimicrobial sensitivity pattern of the pathogen has not changed’. UnfortuClinical Microbiology and Infection, Muzembo et al. conduct asystematic review and meta-analysis of the evidence accumulated over more than 15 yearson the performance of these tests [Fortunately, a handful of immunochromatographic lateral-flow assays, referred toas rapid diagnostic tests (RDTs), are available on the market today. In this issue ofse tests . ThemetV. cholerae. Recent use ofRDTs in both endemic and outbreak situations, including Yemen where more than 400 000RDTs (roughly 25% of the suspected cases reported with a 40% positivity rate) have beenused since 2018, make it clear that decentralized RDT use can be possible even in someof the most challenging field conditions . Although broad use of RDTs is possible, a number of critical policy andscientific steps are needed to move towards rational widespread use of these diagnostictools.If used appropriately, RDTs can help us transition from making epidemiologicalinference and public health decisions on counts of acute watery diarrhoea cases tocounts of acute water diarrhoea caused by Rigorous standards for cholera RDTs in terms of both performance and qualitycontrol are needed to ensure that only high-quality tests are used and to allow forappropriate interpretation of results at the individual and population-levels. Overthe past decade, manufacturers of commonly used RDTs have made significant and oftenundocumented changes to their devices without appropriate communication to endusers, eroding trust and making it hard to compare results of validation studiesconducted in different time periods . In2017Although new guidance from WHO on RDT use has not been released, interimguidance on RDT use from the WHO-led GTFCC in 2015 stated that an RDT positive issufficient to launch a cholera alert but not a confirmed outbreak, which requiresculture or PCR results . This inOne-size-fits-all recommendations will be challenging, if not impossible, todevelop, but we urge simple guidance that clinical staff can implement andunderstand. For example, testing all of the first five suspected cases each day ateach facility then sampling every tenth suspected case after that. This type ofguidance can help ministries of health to plan for the procurement of RDTs,integrate RDT results into routine surveillance system reports  and allow for a more refined picture of cholera locally andglobally. Although it appears that testing of samples after a multi-hour enrichmentstep improves specificity, this comes with operational shortcomings (i.e. it makesthe rapid test no longer rapid) and guidance on when to use tests directly on stoolor after enrichment, perhaps depending on context (e.g. in district laboratoriesrather than in the cholera treatment units), would further improve adoption ofRDTs.V. cholerae strain, co-circulating pathogen, ‘randomchance’, or simply an insensitive reference standard) more appropriateguidance can be crafted, especially that relating to the detection of outbreaks. Asshown by Muzembo et al., the heterogeneity of performance estimates across studieshas been extremely large across settings and tests. A well-controlled head-to-headfield comparison of quality-assured RDTs against both PCR and culture would greatlyimprove our ability to understand the real differences between assays and theiractual field performance when conditions are close-to-ideal.Our understanding of the current generation of RDTs is far from perfect.Almost all field evaluations of RDTs have reported false-positive results, but thishas not been well replicated in laboratory studies . Making V.cholerae O1 and the fact that in many places even after choleratransmission is largely eliminated, we may expect to see diarrhoea caused bynon-toxigenic V. cholerae O1, which are not typically involved inlarge epidemics but will test positive with current RDTs [V. cholerae strains,more work is needed to understand whether the RDTs used could serve as anappropriate medium to temporarily preserve and transport the genetic material neededfor subsequent molecular analyses, as has been done with filter paper [Looking to the future, the current generation of commercially available RDTsmay not be sufficient. As countries progress on their pathway to choleraelimination, the specificity of the current generation of tests (up to 90%) may betoo low. This includes both the analytical specificity in detecting ent RDTs . New raper paper .Culture and PCR both have critical roles to play in the cholera diagnosticslandscape, but RDTs provide an important avenue to decentralize (e.g. expand testingto more primary-care facilities) and refine surveillance systems. These simpletools, while imperfect, can sharpen our view of this disease by distinguishing truecholera from other causes of acute watery diarrhoea. This sharpened view will helpthe global community more efficiently target cholera resources, including vaccines,and make progress towards the goal of ending cholera as a public health threat by2030 and beyond.


Breast cancer (BC) is one of the diseases with the highest female mortality rates in the world and is closely related to breast cancer stem cells (BCSCs). Conventional breast cancer chemotherapy drugs target noncancer stem cells (non-CSCs), while cancer stem cells (CSCs) can still survive, which is an important reason for breast cancer drug resistance and local recurrence or distant metastasis. How to eradicate BCSCs while killing BCs is the key factor to improve the effect, and it is also an important scientific problem to be solved urgently. Therefore, targeted BCSC therapy has become a research hotspot. Interestingly, the emergence of nanotechnology provides a new idea for targeting BCSCs. This study summarizes the current application status of nanomaterials in targeting BCSCs, and attempts to construct a new type of lipid nanoparticle (LNP) that can target BCSCs through mRNA, providing a new idea for the treatment of BC. The International Agency for Research on Cancer (IARC) released the latest global cancer burden data, showing that the number of new cases of breast cancer (BC) worldwide reached 2.26 million in 2020, making it the common form of cancer in the world. BC is a stem cell disease characterized by the existence of cancer cells with stem-like features and tumor-initiating potential. These cells contribute to tumor dissemination and metastasis . Most chBCSCs were first identified and isolated by Al-Hajj  from a pAlthough the treatment of BC has made great progress, the mortality rate of BC still ranks first among female malignant tumors, mainly due to the recurrence and metastasis of BC. Data show that the recurrence and metastasis rate of BC is still as high as 23.4%, which is closely related to BCSCs –10. The + CD24− BCSCs, increase the cell clonogenesis and promote the migration and invasion of cancer cells [Metastasis is a marker of malignant tumors and one of the main causes of death in BC patients and other cancer patients. BCSCs may be the basis of tumor invasion and metastasis and may promote BC metastasis when special dry markers are highly expressed or under the action of the tumor microenvironment . It is noteworthy that the proliferation and metastasis of BCSCs can also be directly regulated by chemokines, transcription factors and proteins. Studies have shown that the chemokine receptor CXCR3B is significantly upregulated in BCSC subpopulations. Ectopic expression of CXCR3B can increase ALDH activity or the number of CD44er cells . Other ser cells .+ cell sets have higher tolerance to chemotherapy, because these cells have the characteristics of BCSCs and are resistant to conventional chemotherapy [+ led to activation of P62-related NRF2 in CD44+ BCSCs, which further promoted the drug resistance of CD44+ BCSCs. In addition, the increased expression of ATP-binding cassette (ABC) family transporters in BCSCs may lead to multidrug resistance in tumors [Drug resistance is a long-term problem in the treatment of BC. BCSCs are resistant to chemotherapy and radiotherapy, and conventional chemoradiotherapy can only target BCSCs with active proliferation, but has no killing effect on BCSCs in the resting state. The retention of BCSCs may increase the pool of CSCs in tumors. BCSCs themselves have inherent resistance to neoadjuvant chemotherapy . Studiesotherapy . Ryoo etotherapy  found thn tumors . The pron tumors . Diedn en tumors  found ran tumors  reportedIn recent years, nanomaterials have been popularized in the application of tumor therapy due to their special size effect and their ability to target tumor sites through the enhanced permeability and retention (EPR) effect. Nanomaterials can effectively improve the problems of poor bioavailability, low specificity and systemic toxicity of traditional drugs and achieve precise and efficient drug delivery at tumor sites.Surgical resection is a common and effective method in the treatment of BC , 25. TheIn recent years, various nanoparticles (NPs) have been discovered and synthesized that can selectively target tumor cells without causing any harm to healthy cells or organs. Therefore, NPS-mediated targeted drug delivery systems (DDSs) have become a promising technique for the treatment of BC. Many agents targeting BCSCs have been proposed by researchers, such as targeting BCSC surface markers, inhibiting BCSC-dependent signaling pathways, interfering with BCSC differentiation, targeting metabolism in BCSCs and targeting the breast tumor microenvironment. However, due to poor water solubility, short cycle time, instability and off-target effects, drugs fail in clinical application. BCSC-targeted drug delivery systems can specifically deliver anti-BCSCs drugs to BCSCs without off-target effects by improving bioavailability. In addition, nanodelivery systems can further enhance BCSCs targeting by surface modification of suitable ligands that interact with overexpressed receptors on the BCSCs surface. The delivery strategies of the nanodelivery system for BCSCs mainly include: delivery of anti-BCSC drugs to tumors; combined delivery of chemotherapy drugs and anti-BCSC drugs to the tumor; active targeted delivery of anti-BCSC agents . Wedelol3O4 nanoparticles and polymer composite nanoparticles, play different roles, and the mediated effect of the field is an important mechanism to kill tumor cells and CSCs. The specific process is under the condition of near-infrared irradiation, between tumor cells and cells residing within nanometer carriers. On the one hand, NIR light is absorbed and efficiently converted into heat energy, which can be stored locally to kill tumor cells [At present, researchers pay more attention to nanocarriers, including polymer nanoparticles (PNPs), liposomes and micelles . The advor cells , 40. On or cells .Table 1CCurrently, polymer nanoparticles are widely used in drug delivery systems, among which PNPs are one of the most recognized nanoparticles and the simplest soft material that can be used as a nanomedicine . AnticanLiposomes are colloidal nanocarriers composed of an amphiphilic phospholipid bilayer, which can be loaded with hydrophilic and hydrophobic drugs. Liposomes have the advantages of biocompatibility, easy surface modification and long circulation time in blood, making them ideal nanocarriers for anti-BCSC treatment. Gemcitabine (GEM)-coated hyaluronic acid-coupled liposomes significantly enhance the cytotoxicity, anti-migration, and anti-colony formation of GEM by targeting CD44 expressed on BCSCs, showing promise for the treatment of breast cancer in vitro and in xenograft models targeting BCSC . The gre+ antibody functionalized PLGA-co-PEG polymer micelles have been used in breast cancer cell line therapy. The results show that encapsulation of PTX into the targeted PLGA-co-PEG micelles increases the sensitivity of BCSCs to PTX [Micelles have a hydrophobic core and dozens of nanosize particles on the surface of a water meter and are usually used as carriers of hydrophobic drugs. Polymer micelles are derived from self-assembled amphiphilic block copolymers. These colloidal particles polymerize with hydrophilic and hydrophobic components, providing a platform for a variety of modifications to improve targeting efficiency. Amphiphilic block copolymers are usually composed of two or three blocks, of which PEG is the most common hydrophilic block in the copolymer structure. Polymer micelles have become popular drug carriers for anticancer therapy due to their uniformity, small size and extended circulation time . Paclitas to PTX . Zhang es to PTX  demonstrLNPs are essentially tiny lipid vesicles, typically assembled from synthetic lipids and other molecules such as cholesterol, phospholipids and polyethylene glycol. The core component, synthetic lipids, is usually composed of a hydrophilic head structure, a hydrophobic lipid tail structure and a linker linking the head and tail. LNPs are widely used in various fields because of their targeting function, such as targeting the precise site of radiotherapy and chemotherapy, and mixing drugs for precise delivery. LNPs are biocompatible vehicles of phospholipid monolayer, structures that can wrap mRNA in lipid nuclei and avoid degradation. The latest study found that mRNA drugs delivered by LNPs may become an effective means for the treatment of LAM, a serious lung disease . Hence, Nucleic acid drugs include antisense nucleic acid (ASO), small interfering RNA (siRNA), microRNA (miRNA), small living RNA (slRNA), messenger RNA (mRNA) aptamer, ribozyme and ARC. As a form of gene therapy, nucleic acid drugs can be made into new drugs simply by rearranging the sequences of A, G, C and T(U). Nucleic acid drugs can target molecules that cannot be targeted by chemical drugs or antibody drugs (such as mRNA and miRNA), which is expected to produce breakthroughs in diseases where traditional drugs have poor efficacy . Due to Nucleic acid drug development also faces three major problems. First, nucleic acid molecules in the body are not stable, nucleic acid drug molecules, regardless of their base composition, structure and endogenous nucleic acid molecules, are vulnerable to all kinds of nucleic acid enzyme degradation, and their RNA—phosphate skeleton structure is electronegative, polar, and completely unable to meet the basic conditions of drug use , 59. TheRNAi (RNA interference) is an important method of targeted therapy, but RNAi transfection efficiency is low, and RNA is easily degraded by intracellular nucleases . The nanSome companies have very similar lipid ratios for nucleic acid delivery systems, with 50% ionizable cationic lipids in LNPs, 10% neutral lipids and 38–40% cholesterol. LNPs mimic membrane-forming cells and are only approximately 100 nm in diameter, consisting of neutral lipids, cationic lipids, cholesterol and PEG lipids. The specific molecular composition of LNPs is shown in Fig. Cationic lipids combined with negatively charged mRNA can efficiently package nucleic acid drugs, while providing positive charge, and negatively charged mRNA compound, cations escape, mRNA transfection in vivo, and ionized lipids have PH sensitivity.Auxiliary lipids: can stabilize particles, destroy the stability of the contents and improve the efficiency of nucleic acid delivery.Cholesterol: stabilizes the LNP structure, regulates membrane fluidity and improves particle stability.Pegylated phospholipids: improve particle stability, reduce particle binding with plasma protein in vivo and prolong systemic circulation time.Stabilizers sucrose or trehalose can improve the stability of LNP and mRNA vaccines and prevent excessive lipid viscosity.For the past few years, increasing interest has been seen in using the LNPs to deliver mRNA therapeutics. The in vitro synthetic mRNA has the potential to produce therapeutically relevant proteins “in vivo” to control and treat a broad spectrum of diseases, including AIDS, rare diseases, cancer, and coronavirus. A research team developed a lung-targeted LNP that specifically delivered the mRNA encoding the normal Tsc2 gene to TSC2-deficient TTJ cells for the treatment of lymphangitic pulmonary fibroids (LAM) caused by Tsc2 gene mutation . Yulia RCD44 is specifically overexpressed in BCSCs and is a specific marker for BCSCs . P245, aCurrently, there are still many problems in the targeted application of novel LNPs to BCSCs. Several studies have been able to combine the CIRSP-Cas9 gene editing system with LNPs for targeted delivery to the lungs, so we can apply gene editing technology to BC and combine gene editing technology with new LNP systems. It is possible to develop an LNP delivery system targeting BCSCs, and complete technological transformation to transform killer tumor cells into a targeted treatment of BCSCs, to reduce the recurrence and metastasis of BC. The innovation of this paper lies in the novel LNP system we constructed, which carries mRNA and adds fluorescent dye to trace, providing a new idea for the early identification and targeted treatment of BCSCs.


Animal behavior, from simple to complex, is dependent on the faithful wiring of neurons into functional neural circuits. Neural circuits undergo dramatic experience-dependent remodeling during brief developmental windows called critical periods. Environmental experience during critical periods of plasticity produces sustained changes to circuit function and behavior. Precocious critical period closure is linked to autism spectrum disorders, whereas extended synaptic remodeling is thought to underlie circuit dysfunction in schizophrenia. Thus, resolving the mechanisms that instruct critical period timing is important to our understanding of neurodevelopmental disorders. Control of critical period timing is modulated by neuron-intrinsic cues, yet recent data suggest that some determinants are derived from neighboring glial cells . As glia make up 50% of the human brain, understanding how these diverse cells communicate with neurons and with each other to sculpt neural plasticity, especially during specialized critical periods, is essential to our fundamental understanding of circuit development and maintenance. This structural remodeling resulted in sustained changes to visual acuity. Importantly, ocular dominance plasticity was limited to a narrow developmental window  around eye opening, after which visual experience had limited impact of circuit structure/function  and other extracellular matrix components that encase the cell bodies and proximate neurites of PV cells. CSPGs are known molecular brakes that prevent remodeling and repair of the injured central nervous system CNS, . In deveWhile much of the critical period field has focused on neuronal mechanisms of critical period timing, such as circuit inhibition, neurons do not function in isolation. A prominent proportion of brain mass is composed of non-neuronal cells called glia, including microglia, as well as the macroglia (astrocytes and oligodendrocytes). Changes in glial development strongly impact neural circuit function, from relatively simple invertebrates through humans . In thisIn development, neurons make more synapses than they ultimately require. During critical periods of plasticity, ectopic synaptic connections are then pruned in an activity-dependent manner, a process that is heavily regulated by all glial types . MicroglIn a series of landmark studies from the Barres and Stevens labs, the authors demonstrated that neuronal activity drives microglial-refinement of retinogeniculate synapses during an early postnatal “pruning period” . In mammWhile the pruning period peaks prior to eye opening, and is thus outside the context of the classic visual critical period defined by Hubel and Weisel . The ECM makes up 20% of the volume of the human brain, serving many diverse roles including cell signaling, barrier functions, and structural support . Glial cfunction . More ret stages . Thus, mBeyond synaptic remodeling, glial cells can both mediate and suppress larger-scale structural remodeling of neurons during developmental critical periods. Oligodendrocytes are the myelinating glia of the CNS, iteratively wrapping axons to form the myelin sheath, an insulating membrane required for rapid action potential propagation. Although oligodendrocytes derive from oligodendrocyte precursor cells (OPCs), OPCs are abundant in the CNS well into adulthood , and incwithin critical periods, glia can non-autonomously regulate critical period timing. These studies shed new light on classic mediators of critical period closure  and unveil new pathways that warrant further investigation  resulted in an ocular dominance shift that is not observed in wildtype controls; importantly, the Nogo-A mutant mice phenocopy this result. Together, these data indicate that ocular dominance plasticity is extended beyond development in the absence of NgR/Nogo-A signaling between neurons and myelinating oligodendrocytes  shape both the expression and timing of neural plasticity. In other words, neurons may be the musicians, but glia may very well be in the conductor’s seat.SA and EH conceived the topic. SA and JS co-wrote the manuscript. JS generated the associated figure. All authors read and approved the manuscript before submission.


Guillain-Barré syndrome (GBS) is a rare autoimmune disorder of the peripheral nervous system that causes progressive weakness and sensory disturbances, usually following an infection or immunization. It has been associated with multiple causes, including bacterial and viral infections. Hepatitis E virus (HEV) is a common cause of acute viral hepatitis that can rarely develop neurological complications. We report a case of a 72-year-old man who developed GBS secondary to an acute HEV infection. He presented with numbness and weakness of the lower limbs that rapidly evolved into respiratory failure requiring mechanical ventilation and intravenous immunoglobulin therapy (IVIg). This case adds to the literature on the association between HEV infection and GBS and the importance of early detection of this rapidly progressive condition. Guillain-Barré syndrome (GBS) is a rare disorder of the peripheral nervous system characterized by progressive weakness and sensory disturbances, usually following an infection or immunization . HepatitA 72-year-old male presented to the emergency department with symmetrical numbness and weakness in both lower limbs. He experienced difficulty walking, which began upon awakening and rapidly progressed to involve both upper limbs symmetrically. He had been discharged the day prior after an elective cataract surgery. The patient's past medical history was relevant for hypertension, type 2 diabetes mellitus with no known microvascular complications, and hypercholesterolemia, and the patient was on Atorvastatin, Azilsartan, Chlorthalidone, Metformin, and Dapagliflozin.Neurological examination revealed a relatively symmetric paraparesis with grade 4/5 strength on lower limbs and diminished sensation in a stocking-glove distribution. Upper limb strength was unaltered. There were no changes in cranial nerves and no signs of meningeal irritation. The remaining physical examination was unremarkable.Laboratory tests showed elevated liver enzymes, and positive serology for HEV with a high viral load Table . HepatitThe patient was admitted to the ward; however, he rapidly deteriorated with worsening quadriparesis, hyporeflexia, dysphagia, and dyspnea. A lumbar puncture was performed, showing albuminocytologic dissociation with increased proteins and a normal cell count Table . ElectroThe patient received a five-day course of IVIg, which resulted in gradual improvement of his neurological status. He was successfully extubated on day 14 and transferred to the neurology ward. Liver enzymes normalized with supportive care, and liver function was preserved throughout the illness. At discharge, the patient maintained areflexia with mild residual weakness in the proximal muscles of the lower and upper limbs and no changes in sensation. He was referred to a rehabilitation clinic for further recovery.This case illustrates a rare but serious complication of acute HEV infection, namely, GBS . GBS is HEV infection is endemic in many developing countries, where it is transmitted mainly through the fecal-oral route via contaminated water or food. However, HEV infection is also increasingly recognized in developed countries, where it is transmitted mainly through zoonotic sources, such as pigs and wild boars . HEV infThe diagnosis of HEV-associated GBS relies on the detection of HEV RNA or antibodies in serum or cerebrospinal fluid (CSF), along with the clinical, laboratory, and electrophysiological features of GBS ,4. CSF eThe treatment of HEV-associated GBS is similar to that of idiopathic GBS, consisting of immunomodulatory therapies, such as IVIg or plasma exchange. The prognosis of HEV-associated GBS is generally good, with most patients recovering fully or partially within six months. However, some patients may have residual neurological deficits or require prolonged mechanical ventilation -6.We report a rare case of GBS secondary to an acute HEV infection, which presented with severe neurological and hepatic manifestations. The patient responded well to IVIg therapy and supportive care and had a favorable outcome. This case underscores the possibility of GBS as a complication of various infections and the need for clinicians to pay attention to rapidly worsening signs and symptoms that may indicate this condition. Early diagnosis and treatment of GBS can prevent life-threatening complications and improve prognosis. This case also adds to the growing literature on the association between HEV infection and various neurological disorders, which may have important implications for the epidemiology, pathogenesis, and management of these conditions.


Many of the bioinformatics tools currently used to analyze EV cargo rely on third-party annotations. Recently, analysis of unannotated expressed RNAs has become of interest, since these may provide complementary information to traditional annotated biomarkers or may help refine biological signatures used in machine learning by including unknown regions. Here we perform a comparative analysis of annotation-free and classical read-summarization tools for the analysis of RNA sequencing data generated for EVs isolated from persons with amyotrophic lateral sclerosis (ALS) and healthy donors. Differential expression analysis and digital-droplet PCR validation of unannotated RNAs also confirmed their existence and demonstrates the usefulness of including such potential biomarkers in transcriptome analysis. We show that find-then-annotate methods perform similarly to standard tools for the analysis of known features, and can also identify unannotated expressed RNAs, two of which were validated as overexpressed in ALS samples. We demonstrate that these tools can therefore be used for a stand-alone analysis or easily integrated into current workflows and may be useful for re-analysis as annotations can be integrated  Liquid biopsy is a broad term used to describe the collection of biological fluids, such as blood, urine, and saliva, to identify biomarkers associated with a specific disease . This apEVs can be categorized into two different classes: exosomes and ectosomes . These sde novo RNA discovery; however, these tools frequently require some form of annotation for differential expression analysis. Most of these tools apply some form of successive alignment steps to various annotations, similar to exceRpt. Alignment-free methods have recently gained significant popularity in genomics and have been adapted to sRNA-Seq with the development of DEUS, an R package for small RNA profiling that is based on the Differential Expression of Unique Sequences  was used for these analyses . Plasma https://github.com/acri-nb/derfinder-pipe.We performed the alignment of our small RNA seq data  with bowA 12-sample small RNA-seq experiment was simulated using the polyester package for R (version 1.20.0) . Reads wAmplification and validation of unannotated RNA sequences were performed similarly to other small RNA (miRNA) as previously described  using reAll relevant sequencing data used in these experiments were deposited to the Gene Expression Omnibus (GEO) with the accession number GSE183942.We first chose to test the applicability of annotation-agnostic tools for RNA sequencing (RNA-seq) data using a simulated dataset. RNA-seq reads were simulated using the polyester package for R before analysis using derfinder, srnadiff, ShortStack or two popular annotation-based quantification methods: HTSeq and featureCounts. Count matrices for specific regions detected with derfinder were generated using the regionMatrix function. Regions in this matrix were then assigned to known features using the GenomicRanges package. Results from derfinder, ShortStack and srnadiff were then compared to those from featureCounts and HTSeq. Feature-wise counts generated by annotation-agnostic methods were highly correlated to those from featureCounts and those from HTSeq for all 12 simulated samples . Out of We were interested in testing the applicability of annotation-agnostic tools for the analysis of small RNA-seq data obtained from liquid biopsy material, namely, EVs. We therefore sought to compare results from annotation-free methods to those from other standard tools that summarize reads to feature-level using real RNA-seq data from EVs, which was previously reported in Furthermore, when quantifying total RNA, the relative abundances of different RNA species such as mRNA, miRNA and lncRNA did not differ substantially between both strategies, with most reads aligning to miRNA, mRNA or lncRNA, which is typical of extracellular RNA sequencing data  Figure . miRNA qAnother frequently used method for analyzing sequencing data in EV genomics consists of aligning reads directly to known transcript annotations, such as miRbase. This was the original approach used for miRNA analysis of the published sample data . We compp-value and rank-correlated. This analysis showed a good (>0.6) correlation of gene ontology (GO) enrichment terms among all tools  common features as significantly differentially expressed, while 163  features were detected by two or more tools . Afterwall tools . The samll tools . Gene-seWe then generated an R script that outputs expressed regions detected by derfinder and Shortstack with several supplementary columns describing overlaps with known features in the provided annotation. When run agnostically and considering only expressed regions rather than genes, 2,672, 2,737, and 2,473 expressed regions were identified by derfinder, ShortStack, and srnadiff after low-count filtering, respectively. Regions identified by all three tools were chosen for subsequent validation by qPCR. For this analysis, alpha was set to 0.1 to maximize the number of candidate biomarker regions.Unannotated expressed regions were mapped to human and exogenous genomes to omit possible confounding sequences, as small RNAs often also map to exogenous organisms. Using BLAST, 56% of unannotated expressed regions could also be mapped to exogenous genomes (not shown). These regions were excluded from downstream validation. Following these analyses, three targets were chosen for subsequent validation by reverse transcription digital droplet PCR, two of which could be successfully amplified. Both RNAs were highly abundant, more than 20 nucleotides in length, and significantly differentially expressed in ALS patient EVs compared to EVs from healthy donors with all three tools . Fifteende novo biomarker discovery and machine learning, especially for liquid biopsies. Thus, in some cases, it may be worthwhile to include expressed RNAs from loci usually considered uninformative, especially with supervised learning algorithms, where the end goal may not necessarily be maximizing interpretability. The field of liquid biopsy research shares many of the challenges of sRNA-Seq analyses since small RNA species are often the most abundantly reported class of RNAs packaged into EVs. In addition, the majority of mRNA and lncRNA sequences detected in EVs are present as fragments of these large RNA species. These challenges are further complicated for EV analyses due to the complex and variable nature of sequencing data for EVs that is caused by multiple cell types contributing to the overall circulating EV pool in most biofluids. It is therefore of prime importance that the bioinformatic methods available for these types of analyses be as accurate, accessible, and complete as possible. Several tools designed specifically for miRNA analysis have been developed (The use of annotation-based approaches, such as direct alignment to an annotation database or application of read-summarization software, for RNA-sequencing data analysis, has many advantages. These approaches quantify read counts within the scope of known high-quality annotations, which increases the interpretability of the data generated by RNA-sequencing experiments. It may also help in avoiding the interpretation of repetitive or otherwise problematic loci; however, some information is lost when restricting analyses to known regions. Post-transcriptional processing of RNA species and incomplete annotations may lead to expressed regions that may not be identified with annotation-based approaches. Reads mapping to regions outside of known annotations are perhaps not as interpretable as those aligning to genomic features; however, they could be valuable in eveloped , but feweveloped . Herein,Using biological data from p-value rank correlation among approaches, indicating that enriched GO terms are likely consistent, regardless of the choice of method. Pathway analysis is fairly correlated among derfinder, HTSeq, featureCounts, and srnadiff; however, pathway enrichment results from ShortStack do not correlate with those from other tools. This indicates that the conclusions drawn from results generated by annotation-agnostic methods generally resemble those from other approaches if using GO annotations, but may show substantial differences depending on which database is queried, especially for pathway enrichment databases. We also observed that srnadiff identifies more significantly differentially expressed regions than other tools when using built-in statistical methods. Therefore, we suggest that anyone considering the analysis of EV data with annotation-agnostic approaches make use of the output of multiple tools, and take great care in the choice of alignment and quantification parameters.Gene-ontology analysis of results from each tool indicates good de novo expressed regions is advantageous for small RNA, as these RNAs tend to be challenging to annotate due to ambiguity in feature start and end coordinates (Here we assigned annotations using a single annotation file from GENCODE, which contains protein coding, miRNAs, lncRNAs, tRNAs, and other non-coding RNAs; however, it is possible to provide a custom annotation file as input for the method, such as piRNAs at pirBase  or tRNA-rdinates . Neverthad hoc basis for the targets chosen in the context of this comparative study, this should be done systematically, as is currently implemented in the exeRpt pipeline (Validation of our data confirmed two unannotated regions as potential biomarkers for ALS, as these were highly over-expressed in ALS samples compared to healthy donors. Caution must be taken when choosing candidate targets for validation, as these may align to recent annotation releases and also to exogenous species, especially for short sequences. Bacterial RNA may be detected in circulation, and while we have verified this on an pipeline . In addi


The burden of cancer is increasing globally. Several challenges facing its mainstream treatment approaches have formed the basis for the development of targeted delivery systems to carry and distribute anti-cancer payloads to their defined targets. This site-specific delivery of drug molecules and gene payloads to selectively target druggable biomarkers aimed at inducing cell death while sparing normal cells is the principal goal for cancer therapy. An important advantage of a delivery vector either viral or non-viral is the cumulative ability to penetrate the haphazardly arranged and immunosuppressive tumour microenvironment of solid tumours and or withstand antibody-mediated immune response. Biotechnological approaches incorporating rational protein engineering for the development of targeted delivery systems which may serve as vehicles for packaging and distribution of anti-cancer agents to selectively target and kill cancer cells are highly desired. Over the years, these chemically and genetically modified delivery systems have aimed at distribution and selective accumulation of drug molecules at receptor sites resulting in constant maintenance of high drug bioavailability for effective anti-tumour activity. In this review, we highlighted the state-of-the art viral and non-viral drug and gene delivery systems and those under developments focusing on cancer therapy. Cancer is a major public health concern and the leading cause of disease-related mortality, with an estimated 10 million deaths worldwide in 2020 . In thisThese emerging delivery systems are designed to transport predetermined quantities of drugs or genes to specific cell receptor sites for selective uptake into diseased cells . Drug deAside from a stable homeostatic environment, other essential factors influencing the functioning of drug or gene delivery systems include molecular size, chemical composition, the bioconjugation approach for attaching payload to the vector stability in biological systems and immune mimicry , 9. The While platforms incorporating the delivery of a drug or therapeutic gene in the context of immune-mediated therapy using delivery vectors are limited, previous attempts to incorporate cancer gene therapy and immunotherapy have yielded positive outcomes –12. ThesDrug delivery is a process which includes the administration, distribution and engagement of the drug and drug molecules with their target aimed at achieving therapeutic effects at site of action and or systemic effects. The term ‘drug’ may represent biologics, peptides, nucleic acids, transgenes or small molecules, among others. Targeted approaches for the delivery of therapeutics to cancer cells typically employ systemic administration or loco-regional delivery targeted at localising the payload at the target site while maintaining steady bioavailability, as illustrated in Fig. Contrary to the dependence on the EPR effects by non-viral drug delivery systems for their mechanisms of action, the design of viral vector purportedly aims at shielding from neutralising antibodies, maintenance of optimal effective dose and stable expression of transgenes independent of the route of administration , 22. Thi(DDS) is to modify the pharmacokinetics of a therapeutic molecule by packaging to control its targeted release and accumulation. Their categorization can be according to different features including size, chemical composition, method of administration, target organ of interest and type of material used. Based on delivering biomaterials, drug (gene) delivery systems may be grouped into two: viral and non-viral systems. The latter includes the use of uniquely identified proteins and nanoscale particles that are engineered to serve as vehicles for drug delivery -N,N,N-triethylammonium (DOTMA) and 1,2-dioleoyl-3-trimethylammoniopropane (DOTAP) which are characterised by having a positive charge (cationic) , 125. ThDue to their versatile structure, biocompatibility, non-immunogenicity, biodegradability and lack of toxicity, lipid-based systems are considered a reliable delivery system for either cytotoxic drugs or genes . They ar®, the first drug delivery system based on a PEGylated liposome technology used to treat AIDS-related Kaposi’s sarcoma, breast cancer, ovarian cancer and other solid tumours. Doxil contains encapsulated doxorubicin hydrochloride, an anti-cancer agent that induces apoptosis in cancer cells by blocking topoisomerase IIα generating oxidative DNA damage that blocks cancer cell division and growth. When used in a conventional manner, doxorubicin will generate cardiotoxic effects, and it has been shown that encapsulated doxorubicin in liposomes helps to overcome the treatment-associated toxicity, showing reduced cardiac toxicity [Specific to cancer, the encapsulation of lipophilic or hydrophilic anti-cancer drugs has been very convenient for the integration of chemotherapeutics into liposomal preparations, as mentioned and perfectly summarised in a recently published review by Yingchoncharoen et al. . One exatoxicity , 130.On the other hand, lipoplexes have also been successfully studied for in vitro and in vivo delivery of miR-29b microRNA to the cytosol of cells for the treatment of lung cancer , for tarThe next generation of lipid nanoparticles include solid lipid nanoparticles (SLNs), nanostructured lipid carriers (NLCs) and cationic lipid-nucleic acid complexes; these have been on the scope from the late 90 s as novel types of gene/drug delivery vectors due to their enhanced physical stability that provides more effectiveness and safety. SLNs are made of physiological lipids dispersed in water or an aqueous surfactant solution, and their size ranges between 50 and 1 μm. Because of their colloidal dimensions, SLNs have several advantages, including improved physical stability and biocompatibility, controlled particle size, prolonged release, easy scale-up and manufacturing and better formulation stability through freeze-drying and reconstitution processes , 138. Ho50 in vitro and inhibiting tumour rates in vivo. This was accomplished by loading NLCs with either the chemotherapeutic drugs alone, in combination with a low dose of an adjuvant, or by conjugating the drug-loaded NLCs with an active targeting moiety. For example, NLCs were used in combination with docetaxel, a drug approved for the treatment of several types of cancer such as breast cancer, non-small cell lung cancer, advanced stomach cancer, head and neck cancer and metastatic prostate cancer; when administering the DTX-NLC in murine melanoma (B16 cell line), the IC50 of the complex was reached with 0.47 μg/ml of treatment, compared to the need of 0.96 μg/ml needed to reach an IC50 when administering the free drug; the same complex was used with hepatocellular carcinoma (HepG2), Pulmonary adenocarcinoma (A549) and ovarian carcinoma (SKOV3) cell lines [The use of NLCs as carriers for chemotherapeutic agents received special attention and demonstrated their remarkable ability to boost the therapeutic effect of some drugs across a wide range of malignancies, enhancing the ICll lines , 142. NLll lines –148. Sinll lines –153. Forll lines .The so-called polyplexes are formed via electrostatic interactions when the negative charges from nucleic acids are complexed with the positive charges of cationic polymers, allowing the delivery of viable DNA or drugs to cells . Due to Another approach of polyplexes was used for efficiently generating reactive oxygen species that helped in endosomal escape of siRNA that was later internalised by cancer cells in vitro and inhibited tumour growth in two xenograft murine models by reducing RRM2 (ribonucleotide reductase subunit M2) expression in cancer which consequently reduced cell proliferation and enhanced cancer cell apoptosis . SeveralPolymeric micelles are auto-assembled core–shell structures formed by different copolymers in the presence of an aqueous system. They are able to hold medications inside their hydrophobic core, whilst the hydrophilic shell gives support and stability to the hydrophobic core and can carry molecules such as DNA or siRNA . Their sDendrimers are spherical tree-like branched synthetic macromolecules possessing unique structural and topological features and are built around a linear polymeric core. They show biomolecule-like properties, a high degreThese cationic PAMAM dendrimers possess ane of physicochemical customization and low polydispersity . DendrimDendrimers have been created in a variety of chemically distinct forms such as polyamidoamine (PAMAM) dendrimers, arborols, poly ether dendrimers, phenylacetylene dendrimers and poly(propylene imine) PPI-dendrimers. Their ability to encapsulate and bind drugs or molecules is continuously explored to enhance solubility, sustain the controlled release of the before-mentioned molecules and deliver drugs into specific sites of the body. Apart from acting as soluble enhancers, dendrimers show additional properties such as non-immunogenicity, spherical structure, biocompatibility, proper biodegradation, minimum non-specific blood-protein binding and the allowance of a regulated drug release, making them suitable as valuable delivery systems for either genes or drugs; for all these reasons, their novelty as a new class of vectors for nanomedicine has been extensively researched in the last couple of years , 183.PAMAM are the most popular dendrimers used for drug delivery systems. They were first reported by Tomalia et al. in 1985, where they stated that dendrimers have “reactive end groups”, consisting of amines and amides that allow for controlled molecular weight building, controlled topology or branching and versatility in the design according to the modification of the terminal end . These cIn cancer therapy, PAMAM dendrimers have been used as efficient tools to deliver chemotherapy agents such as doxorubicin , 189 metPoly(propylene imine) dendrimers (PPI-dendrimers) were first reported by Vogel et al. in 1978. PPI dendrimers have a slightly less polar interior than PAMAM dendrimers, but both have amino terminal functions that make the molecule water soluble. High-generation PPI-dendrimers often have greater DNA complexation than low-generation dendrimers, but they also have increased cytotoxicity and limited manufacturing and purification , 199. AdIn regard to nucleic acid delivery, PPI-dendrimers have been a great improvement for siRNA delivery, since “unprotected” siRNA is prone to rapid degradation and low penetration ability across the plasma membrane . KesharwThere are increasing number of cytotoxic payloads currently undergoing development and evaluation including the high number of already approved anti-cancer drugs for the therapy of solid tumours. Despite these milestones, significant challenges namely non-specific killing of normal cells still persists resulting in poor therapeutic outcome. Indeed, the future of drug (gene) delivery to target polygenic and complex diseases such as cancer is promising. However, the delivery vector must meet strict criteria of safety, target-specific, ability to encapsulate large anti-cancer payload, biocompatibility to discourage activation of immune response against it. Thus far, the field of drug or gene delivery is at the transformative stage whereby breakthroughs are not far from reality.To ensure efficient interaction of chemically conjugated drug delivery vectors and their targets, computational approaches and supercomputing tools employing machine learning methods have been proposed for drug formulations that are currently in active clinical trials . These c. Again, such novel viral vectors with minimal propensity to generate immune responses are essential.For viral vectors, the ablation of the natural binding domain for site-directed retargeting is equally important as it attempts to achieve higher vector yields. Logically, achieving higher viral vectors which are not designed for site-specific targeting becomes a nuisance. However, platforms to integrate these, while conserving viral infectivity and transduction, are lacking. The efficient interaction of therapeutic and diagnostic biomaterials distributed by viral vectors for the penetration of solid tumours as well as overcoming the complex tumour microenvironment is the much-desired gold standard required to harness their potentials as effective gene and drug delivery systems for clinical useOn the other hand, the design of non-viral delivery strategies to aim at overcoming poor tumour transduction by coupling them with cytotoxic ligands is a promising approach. Recent evidence from imaging mass cytometric studies reveals an unequal proportion between distribution of active drug molecules into tumour microenvironment and the delivery of parent drug by non-viral vectors . To over