Inflammatory bowel disease (IBD) is a complex disease characterized by persistent chronic inflammation of the gastrointestinal tract and periodic episodes. Despite the increasing number of related studies, the detailed pathogenesis of IBD has not been elucidated. In recent years, host-microbiota interactions in the pathogenesis of IBD have received extensive attention. Testes-specific protease 50 (TSP50) is a potential risk gene for IBD, but whether it can affect the susceptibility of colitis by regulating the gut microbiome is still unclear. Here, we showed that TSP50 deficiency in neural stem cells (NSCs) aggravated colitis in mice by altering intestinal microbiome. Mechanistically, TSP50 maintained the level of neurotransmitter acetylcholine (ACh) by degrading acetylcholinesterase (AChE), thereby maintaining intestinal mucosa and intestinal microecological homeostasis and reducing the susceptibility to colitis. These findings provide a new perspective on the interaction between host and commensal microbiota, which may be beneficial for developing potential therapeutic strategies for IBD.

Pluripotent stem cells have long been used to produce knockout mice via germline chimera technology. However, aside from the rat, this approach has not been successfully applied to other mammals. Here, we demonstrate that rabbit induced pluripotent stem cells (iPSCs) can be reprogrammed using KLF2, ERAS and PRMT6, enabling them to efficiently colonize embryos. These chimeric embryos can develop into fetuses and newborn rabbits, with iPSCs contributing up to 100 % to certain organs. Notably, female rabbits generated through this method are healthy and transmit the iPSC genome to their offspring with a high efficiency, demonstrating germline chimerism. This advancement establishes a foundation for developing rabbit models of human disease with complex genetic traits.

Mesenchymal stromal cell-derived extracellular vesicles (MSC-EVs) are recognized as a promising strategy for cell-free therapy, however, their therapeutic role in pulmonary fibrosis remains unrevealed. Here, we report the safety and efficacy of MSC-EVs from human umbilical cord (hUCMSC-EVs) evaluated in mouse models and pulmonary fibrosis patients. We established a rigorous system to produce high-quality of hUCMSC-EVs, characterized by miRNA, protein, and metabolite profiles. When administered via nebulization, hUCMSC-EVs predominantly accumulated in murine lungs and ameliorated bleomycin-induced pulmonary fibrosis, with increased survival rate (from 20% to 80%), restored lung volume, and attenuated injury severity accompanied by elevated oxyhemoglobin saturation and improved pulmonary function evaluations. We performed a phase l clinical trial involving twenty-four patients in a randomized, single-blind, and placebo-controlled study to treat pulmonary fibrosis (MR-46-22-004531, ChiCTR2300075466). All participants tolerated the nebulized hUCMSC-EVs well, with no serious adverse events. Patients receiving the combined therapy of nebulized hUCMSC-EVs and routine treatment demonstrated significant improvements in both lung function indices (forced vital capacity and maximal voluntary ventilation) and respiratory health status (as measured by the Saint George's Respiratory Questionnaire and Leicester Cough Questionnaire. Overall, patients upon the additional therapy with nebulized hUCMSC-EVs gained significant benefits compared with those accepted only routine treatment. Remarkably, two patients with advanced post-inflammatory pulmonary fibrosis exhibited clinically significant regression on serial CT scans after hUCMSC-EVs therapy. These findings suggest that nebulized hUCMSC-EVs could be used as a promising therapeutic strategy for treating pulmonary fibrosis diseases.

SARS-CoV-2 entry into host cells is mediated by the spike protein, which drives membrane fusion. While cryo-EM reveals stable prefusion and postfusion conformations of the spike, the transient fusion intermediate states during the fusion process remain poorly understood. Here, we design a near-native viral fusion system that recapitulates SARS-CoV-2 entry and use cryo-electron tomography (cryo-ET) to capture fusion intermediates leading to complete fusion. The spike protein undergoes extensive structural rearrangements, progressing through extended, partially folded, and fully folded intermediates prior to fusion-pore formation, a process that depends on protease cleavage and is inhibited by the WS6 S2 antibody. Upon interaction with ACE2 receptor dimer, spikes cluster at membrane interfaces and following S2' cleavage concurrently transition to postfusion conformations encircling the hemifusion and initial fusion pores in a distinct conical arrangement. S2' cleavage is indispensable for advancing fusion intermediates to the fully folded postfusion state, culminating in membrane integration. Subtomogram averaging reveals that the WS6 S2 antibody binds to the spike's stem-helix, crosslinks and clusters prefusion spikes, as well as inhibits refolding of fusion intermediates. These findings elucidate the entire process of spike-mediated fusion and SARS-CoV-2 entry, highlighting the neutralizing mechanism of S2-targeting antibodies.

This study aims to investigate the acute effect of local allantoin treatment on experimental distraction osteogenesis. A total of 14 adult New Zealand rabbits underwent mandibular distraction under general anesthesia. The control group received saline washes, while the experimental group received local allantoin (1 mg/kg). Computed tomography was used to measure bone mineral density (Hounsfield Units). Stereological analysis of decalcified tissues was performed using the Cavalieri Method, which assessed the volumes of new bone, connective tissue, and capillaries (mm3). Immunohistochemistry employed anti-RUNX2 and anti-BMP2 antibodies. It was found that the volumes of new bone, connective tissue, and capillary were higher in the experimental group than in the control group. Furthermore, anti-RUNX2 and anti-BMP2 activities were more intense in the connective tissue area of the experimental group. Using stereological and immunohistochemical methods, this study has demonstrated that allantoin, known for its antioxidant and anti-inflammatory properties, enhances bone regeneration in distraction osteogenesis. Allantoin contributes to bone healing by stimulating mesenchymal stem cells and enhancing neoangiogenesis.

This study primarily investigates the effect of Liuwei Dihuang Pills on the activation and proliferation of female germline stem cells(FGSCs) in the ovaries and cortex of mice with premature ovarian failure(POF), and how it improves ovarian function. ICR mice were randomly divided into the control group, model group, Liuwei Dihuang Pills group, Liuwei Dihuang Pills double-dose group, and estradiol valerate group. A mouse model of POF was established by intraperitoneal injection of cyclophosphamide. After successful modeling, the mice were treated with Liuwei Dihuang Pills or estradiol valerate for 28 days. Vaginal smears were prepared to observe the estrous cycle and body weight. After the last administration, mice were sacrificed and sampled. Serum levels of estradiol(E_2), follicle-stimulating hormone(FSH), luteinizing hormone(LH), and anti-Müllerian hormone(AMH) were measured by enzyme-linked immunosorbent assay(ELISA). Hematoxylin-eosin(HE) staining was used to observe ovarian morphology and to count follicles at all stages to evaluate ovarian function. Immunohistochemistry was used to detect the expression of mouse vasa homolog(MVH), a marker of ovarian FGSCs. Immunofluorescence staining, using co-labeling of MVH and proliferating cell nuclear antigen(PCNA), was used to detect the expression and localization of specific markers of FGSCs. Western blot was employed to assess the protein expression of MVH, octamer-binding transcription factor 4(Oct4), and PCNA in the ovaries. The results showed that compared with the control group, the model group exhibited disordered estrous cycles, decreased ovarian index, increased atretic follicles, and a reduced number of follicles at all stages. FSH and LH levels were significantly elevated, while AMH and E_2 levels were significantly reduced, indicating the success of the model. After treatment with Liuwei Dihuang Pills or estradiol valerate, hormone levels improved, the number of atretic follicles decreased, and the number of follicles at all stages increased. MVH marker protein and PCNA proliferative protein expression in ovarian tissue also increased. These results suggest that Liuwei Dihuang Pills regulate estrous cycles and hormone disorders in POF mice, promote the proliferation of FGSCs, improve follicular development in POF mice, and enhance ovarian function.

Personalized immunotherapy of pancreatic ductal adenocarcinoma (PDAC) through T-cell mediated targeting of tumor-specific, mutanome-encoded neoepitopes may offer new opportunities to combat this disease, in particular by countering recurrence after primary tumor resection. However, the sensitive and accurate calling of somatic mutations in PDAC tissue samples is compromised by the low tumor cell content. Moreover, the repertoire of immunogenic neoepitopes in PDAC is limited due to the low mutational load of the majority of these tumors.

Myocardial Infarction (MI) is still a leading cause of mortality, and current treatments primarily focus on symptom alleviation and blood flow restoration, with limited capacity for myocardial repair. Exosomes, key mediators of intercellular communication, have demonstrated potential to promote myocardial regeneration but exhibit limited cardiac-targeting efficiency due to rapid accumulation in other organs. To overcome this limitation, we designed targeted nanobubbles (TNBCD81-cRGD) loaded with exosomes derived from adipose-derived stem cells (ADSCs) in this study. These ADSCs were genetically modified through viral transfection to secrete exosomes with high expression of stromal cell-derived factor 1α (SDF-1α), which was upregulated in the infarcted region and promotes stem cell homing via the SDF-1α-CXCR4 axis. The nanobubbles, modified with anti-CD81 antibodies and cRGD, enabled efficient targeting of ischemic myocardium under Low-Intensity Pulsed Ultrasound (LIPUS) irradiation. Our study demonstrated that the combination of targeted nanobubbles, ADSC-derived exosomes with high SDF-1α expression, and LIPUS irradiation enhanced exosome retention in the heart, improved therapeutic efficacy, and promoted myocardial repair. This approach holds potential for advancing exosome-based therapies in myocardial infarction treatment.

Traumatic Brain Injury (TBI) usually leads to substantial mortality and disability among adult populations. Neural progenitor cells (NPCs) transplantation exhibits great potential in TBI treatment. However, the differentiation rate of interneurons is relatively low, largely impeding the therapeutic effects of brain tissue repair.

Despite new therapies for cervical cancer, innovative strategies are essential to overcome drug resistance and high toxicity. The present study focuses on the metabolic profiling of cervical carcinoma using a non-targeted metabolomics approach using liquid chromatography-mass spectrometry. Our study identified over 70 metabolites in cervical tissue samples (both cancerous and adjacent normal) using HILIC and reversed-phase chromatography in the positive and negative ionization modes. Major metabolic alterations included changes in nicotinamide metabolism, ammonia recycling, amino acid metabolism and nucleotide metabolism, in a grade-dependent manner. Compared to normal tissue, HPV-positive tumors showed elevated nicotinamide metabolism, and phosphatidylethanolamine biosynthesis, whereas HPV-negative tumors showed enriched purine and pyrimidine metabolism. We validated our findings by analyzing transcriptomics datasets from the Gene Expression Omnibus database to understand the expression patterns of the underlying genes involved in the dysregulated pathways. We observed that nicotinamide metabolism exhibits significant effects in lower-grade cervical cancers and specific HPV genotypes. We treated cervical cancer cell lines with niacinamide (NAM), an amide form of niacin, to evaluate its therapeutic efficacy. NAM treatment modulated NAD+ metabolism, affecting key players such as CD38, PARP, NAMPT, and SIRT1, promoting apoptosis and inhibiting cell proliferation in cervical cancer cells. Importantly, HPV-positive SiHa cells showed elevated NAD+ metabolism relative to HPV-negative C33A cells, reflecting distinct metabolic adaptations that may influence tumor progression. The study highlights the metabolic shifts in cancer progression and provides insights into NAM's molecular mechanisms and therapeutic potential for precision medicine in cervical cancer.

The rapid increase in emerging contaminants driven by industrialization poses significant threats to both ecosystems and neurodevelopmental health, underscoring the urgent need for sensitive and accurate screening platforms for developmental neurotoxic compounds. Here, we developed a zebrafish-based multi-indicator assessment system to overcome the limitations of conventional behavioral assays. The morphologies of the head, actions of microglial cells, numbers of motor neurons, changes in neuronal activity and behavioral mobility of zebrafish larvae are examined and set as assessment criteria. When validated with 12 known neurotoxicants, our approach significantly improved detection rates compared to previous behavioral screening methods. Specifically, 8 compounds (66.67 %) affected either interocular distance or midbrain area. 10 neurotoxic pollutants (83.33 %) were identified by observing the actions of microglia, while 9 compounds (75 %) showed effects on the patterns of neuronal cell activity and 7 compounds (58.33 %) were identified by the number of motor neurons. Furthermore, a scoring system was established by integrating these indicators, enabling simultaneous hazard identification and risk prioritization. This multi-dimensional assessment approach substantially minimizes the risk of omissions associated with relying on a single indicator. Overall, the sensitivity and accuracy of this method provide an efficient tool for screening and evaluating potential developmental neurotoxic pollutants, thereby supporting the management and risk mitigation of emerging contaminants.

Bones are living tissues that are periodically renewed through the bone remodeling process. It starts when monocyte-macrophage-type cells begin to proliferate and recruit into the bone tissue due to the action of various proteins. The proteins are generated by the body in response to a lack of calcium in the blood or a damaged bone tissue. Then, other chemical signals promote the differentiation of monocyte-macrophage-type cells into osteoclasts. Finally, osteoclasts carry out a process known as bone resorption in which they degrade bone tissue. After, bone resorption a process known as osteogenesis takes place. In osteogenesis, pluripotent mesenchymal stem cells proliferate and recruit in the bone damaged by bone resorption in response to the action of different compounds. Then, pluripotent mesenchymal stem cells differentiate into osteoblasts due to the effect of specific proteins. Finally, osteoblasts generate new bone tissue, completing the bone remodeling process. This review aims to summarize and provide recent findings of the proteins and chemical signals involved in osteoclast and osteoblast proliferation, recruitment and maturation during the bone resorption and osteogenesis processes. The information found about these molecules could help to better understand all the complex mechanisms that directly or indirectly influence the bone remodeling process. In this sense, the mechanisms of different bone disorders and diseases could be elucidated in a better way, leading to the generation of efficient and specific treatments for each of them in the future.

Swine enteric coronavirus-derived disease is an acute, highly contagious and infectious disease that causes symptoms such as diarrhea, vomiting, dehydration and high lethality in suckling piglets, posing a serious threat to the global pig farming industry. The main viral pathogens responsible for the disease include TGEV, PEDV, PDCoV, and SADS-CoV. The investigations of these viruses have been severely impeded by the lack of good in vitro culture systems and small animal models. Intestinal organoids are derived from crypt stem cells, which undergo growth, development and differentiation into structures resembling the intestinal tube morphology, thereby serving as an effective model for studying enteric pathogen‒host interactions in a manner that is as close as possible to in vivo conditions. This review outlines the establishment, regulation, and applications of intestinal organoids, with a particular focus on the advancements in research made possible by swine enteric coronaviruses using this model. The limitations of the intestinal organoids model and potential avenues for future improvement are also discussed. Finally, the findings emphasize the benefits of intestinal organoid models in investigating intestinal pathogen‒host interactions and how they will continue to offer a valuable platform for swine enteric coronavirus research with further developments in intestinal organoid technology.

Rab proteins, a subset of small monomeric GTPases, represent the largest subfamily within the Ras superfamily, which also includes Ras and Rho proteins. Over 70 Rab members have been identified, acting as molecular switches that toggle between an active GTP-bound state and an inactive GDP-bound state. Rab GTPases are crucial for intracellular vesicle formation and membrane trafficking. Stem cells, known for their self-renewal and multipotent differentiation abilities, likely depend on paracrine signaling mechanisms, essential for their application in regenerative medicine. Exosomes, pivotal in cellular paracrine communication, are distinguished by their abundance, biocompatibility, and low immunogenicity. Rab27, a member of the Rab family, is critical in exosome secretion by aiding the targeting of multivesicular endosomes to the cell periphery and their docking at the plasma membrane. Cancer stem cells, a tumor cell subset with unlimited proliferative capacity, can initiate tumor formation. Recent research indicates that Rab27 also influences tumor initiation and progression by modulating exosome secretion. This review summarizes Rab27-mediated exosome secretion regulation and its biological effects on various types of mesenchymal stem cells and cancer stem cells.

CD117 is a cell surface receptor expressed on hematopoietic stem and progenitor cells (HSPCs) and acute myeloid leukemia (AML), and thus CD117-targeting CAR T cells (CART117) can function as both conditioning for hematopoietic stem cell transplant (HSCT) and a therapy for AML. We developed human and mouse CART117 to evaluate the safety and feasibility of targeting CD117 in preclinical mouse models. Human CART117 had potent anti-tumor activity while also mediating significant hematopoietic toxicity in a humanized mouse model. Murine CART117 (mCART117) led to systemic and hematopoietic toxicity without anti-leukemic benefit in immunocompetent C57BL/6 mice. Intriguingly, mCART117 was able to eliminate CD117+ cells in the spleen but not in the BM. Of note, proliferation of BM CD117+ cells in response to lymphodepleting chemotherapy amplified mCART117-mediated systemic toxicity. Alternative lymphodepletion with radiation ameliorated the systemic toxicity of mCART117 but did not improve anti-leukemic efficacy. Immunodeficient mice given mCART117 in the absence of lymphodepletion died from severe pancytopenia, and this effect was recapitulated by Treg depletion in immunocompetent mice. Increasing CD117 expression on AML improved the anti-leukemic efficacy and toxicity profile of mCART117. In conclusion, mCART117 anti-leukemic activity is impaired in immunocompetent mice when CD117 is expressed at physiological levels on AML.

Adenosine Deaminase Severe Combined Immunodeficiency (ADA-SCID) is a monogenic disorder caused by mutations in the ADA gene. Gene therapy utilizing γ-retroviral and lentiviral vector gene addition approaches have shown curative results. We sequenced the ADA transgene in transduced CD3+ T-cells, and in peripheral blood cells from patients treated with autologous CD34+ cells transduced with either a γ-retroviral or lentiviral ADA gene vector to assess transgene mutational profiles. In both CD3+ T-cells and ADA-SCID patients' cells treated with the lentiviral vector, we observed significantly higher occurrences of guanine (G) to adenosine (A) base substitutions than with the γ-retroviral vector. We hypothesized that this G-to-A mutational signature was due to the APOBEC3 cytosine deaminase protein family. By knocking out APOBEC3 genes in HEK239T packaging cells, APOBEC3-mediated mutagenesis decreased by 91.2% along the transgene in CD34+ transduced cells in comparison to CD34+ cells transduced with lentiviral supernatant packaged in parental HEK293T cells.

Inflammatory diseases are associated with immune imbalances and the linkage between immune system and inflammatory mechanisms, particularly under hypoxia, is still incomprehensive. Furthermore, development of a natural agent necessitates, in lack of therapeutics, to prevent thymus receding under hypoxia-induced inflammation. Rat primary thymocytes were exposed to hypoxia (0.5% O2) in the absence or presence of aqueous extract of Ganoderma lucidum. After exposure, redox status and bioenergetic and biochemical variables were evaluated. Hypoxia exposure to thymocytes results in cell growth arreast and death with concurrent surge in reactive oxygen species and decrease in total antioxidant status. Moreover, these hypoxic cells had imbalanced redox homeostasis and bioenergetics with decreased NAD/NADH and increased ADP/ATP ratio. The cells treated with an aqueous extract of G. lucidum rescue cells from death, reverse redox imbalance and changes in bioenergetic markers. Hypoxia exposure enhance pro-inflammatory cytokines; IL6, TNFα, and NFκB in thymocytes which normalizes with the extract treatment. Hypoxic cells displayed higher expression of Nrf2, HIF1α, VEGF in rat thymocytes but downplayed role of erythropeotin. The extract treatment overtakes adverse hypoxia-induced changes on hypoxic markers. Thus, G. lucidum reverses redox imbalance, changes in bio-energetic markers, and displayed strong anti-stress and anti-inflammatory effect.

T and NK cell lymphoma with abnormal CD20 expression is a condition that primarily seen in Asians. Due to the variable expression of CD20, it is difficult to generalize its origin, leading to controversial conclusions about the role of CD20. In this review, we provide an overview of CD20 localization and function in three cell types. In NK/T-cell lymphoma (NKTCL), CD20 was discovered to be confined to B cells, presenting a mature active phenotype and having an inert course. However, in T-cell lymphoma (TCL), variable expression of CD20 antigen on T cells and NK cells correlates with tumor transformation and rituximab efficacy. We highlight the fact that CD20 expression is stage-specific and further classify CD20 antigen assignment, which contributes to understanding the variety of outcome and the function of CD20 during various phases of tumor development.

The intestinal mucosa has evolved to facilitate interactions between the host and the constellation of intestinal microbes, collectively termed the microbiota. A well-orchestrated balance exists in the healthy mucosa where microbes and microbial products first encounter a barrier formed by a single layer of intestinal epithelial cells (IECs). This homeostasis exists at a harsh interface between the highly vascularized mucosa and the anaerobic intestinal lumen. This steep oxygen gradient establishes 'physiological hypoxia' as a central metabolic characteristic of the mucosa. Recently, interest in understanding the dynamic host-microbe interplay has identified microbial metabolites that support host functions at several different levels. Of singular relevance are short-chain fatty acids, particularly butyric acid. Studies have demonstrated that IECs have evolved to benefit from butyrate through a plethora of functions, including energy procurement, metabolism, barrier and wound healing regulation, production of antimicrobial peptides, etc. Butyrate is consumed by differentiated colonic epithelial cells preferentially for energy, creating a distinct butyrate gradient along the intestinal cryp-tvillus axis. The depletion of butyrate and butyrate-producing microbes during active inflammation, termed dysbiosis, promotes disease and attenuates tissue healing responses. Furthermore, in a disease state, the butyrate gradient is disrupted leading to reduced utilization of butyrate and inhibition of proliferation of colonic stem cells. Emerging studies suggest that chemical modifications to butyrate could be useful in targeting select IEC functions for particular benefits to the host. In this review, we consider how butyrate molecular mimicry may play out in the setting of mucosal health and disease and discuss current discoveries on endogenous and synthetic butyrate-like compounds and their pathways.