Objective: Aging is commonly associated with a decline in immune function, hormonal imbalances, insomnia, constipation, erectile dysfunction, and general muscle weakness. To address these complex conditions, our study used probiotic therapy. Methods: We investigated the effects of two spore-forming probiotics, Clostridium butyricum and Bacillus coagulans , on health parameters in 126 men aged 48–64 years through a population-based intervention. Results: This intervention notably improved age-related conditions, particularly erectile function (measured by the Erectile Function Scale Index [EFSI]), bowel movement regularity, and sleep quality. Among 35 blood biochemical parameters, 19 showed significant changes post-treatment, including those related to immune response, hormone levels, inflammation, and lipid metabolism. Some blood parameters associated with probiotic treatment were identified using exploratory machine learning analysis. Conclusions: The results suggest that supplementation with Clostridium butyricum and Bacillus coagulans may be associated with improvements in certain age-related health parameters in older men.

Injectable hydrogels have gained considerable attention as sutureless bioadhesives for minimally invasive wound closure, owing to their ability to adapt to complex wound geometries. However, upon application, these biomaterials can occupy the wound space, potentially hindering critical healing processes such as angiogenesis and granulation tissue formation. To overcome this challenge, we developed a greener synthesis route for the fabrication of alanine-modified cellulose nanofibrils (Ala-CNFs). Compared with traditional cellulose modification strategies, this route eliminates the need for cytotoxic reagents and metal catalysts that are indispensable for conventional approaches. The Ala-CNFs formed a versatile hydrogel via crosslinking with polyvinyl alcohol (PVA) and phenylboronic acid modified cyclodextrin (PBCD) (denoted as Ala-CNFs@PVA/PBCD). Furthermore, the Ala-CNFs@PVA/PBCD hydrogel integrates multiple essential functionalities, including injectability, self-healing, biodegradability, and shape adaptability. Ala-CNFs served a dual function: (i) they enhanced adhesion, enabling the hydrogel to achieve rapid hemostasis in cardiac (38 s) and even femoral artery (34 s) wounds, and (ii) they actively promoted angiogenesis and granulation tissue infiltration within its macroporous matrix, thereby addressing the critical issue of physical barrier posed by conventional injectable adhesives. Consequently, this work not only provides a novel synthesis strategy for functionalized cellulose but also presents a sustainable and multifunctional strategy for advanced wound management, particularly suitable for emergency scenarios such as warfare and earthquakes.

Upon activation, macrophages generate substantial levels of reactive nitrogen species, which can induce alkylating damage in the DNA of intracellular Mycobacterium tuberculosis (Mtb) and thereby restrict bacterial replication. However, the molecular mechanisms by which Mtb repairs such DNA lesions remain poorly understood. Here, we identified genes required for Mtb survival in distinct macrophage subsets using transposon insertion sequencing. Among these, tagA displayed a specialized role in Mtb survival in M1-polarized macrophages, as well as in mice at 4 wk postinfection, a stage when macrophages are biased toward an M1-polarized state. Mechanistically, TagA conferred resistance to the DNA alkylating agent methyl methanesulfonate through its 3-methyladenine (3-MA) excision activity with Glu48 serving as a key catalytic residue for substrate binding. Critically, TagA was found to protect the Mtb genome from alkylation damage caused by nitrosative stress-a hallmark of the M1-polarized macrophage microenvironment. Furthermore, pharmacological inhibition of inducible nitric oxide synthase (iNOS) with S-methylisothiourea sulfate in mice or genetic deletion of nos2a in zebrafish markedly rescued the survival defect of ΔtagA. Together, these findings reveal a previously unappreciated mechanism by which the DNA repair enzyme TagA protects Mtb against 3-MA DNA damage under nitrosative stress, thereby promoting bacterial survival in M1-polarized macrophages and during in vivo infection.

Biomacromolecules based injectable and self-healing hydrogels are promising soft biomaterials with dynamic and reversible features. Yet, it remains a challenge to achieve both high mechanical strength and dynamic behavior. Here, we present a tough dynamic nanocomposite hydrogel (Fe3O4/PDA@OSA-l-Gel) fabricated by incorporating polydopamine coated iron oxide nanoparticles (Fe3O4/PDA NPs) into dynamically crosslinked biomacromolecular networks of oxidized sodium alginate (OSA) and gelatin (Gel) in the presence of borax. The loading of Fe3O4/PDA NPs significantly enhances the mechanical properties of dynamic nanocomposite hydrogel exhibiting excellent injectable and self-healing properties. The synergistic effect of multiple dynamic chemical bonds (e.g., imine bond, borate ester bond) and physical interactions (e.g., electrostatic interaction, hydrogen bond) that enable excellent mechanical properties (3.78 MPa compressive strength, 674.72 J/m3 energy dissipation and 109.1 kJ/m3 toughness). Fe3O4/PDA NPs play the role of killing two birds with one stone, i.e., act as a nano-crosslinker to enhance mechanical properties of the dynamic nanocomposite hydrogel, and as a carrier for sustained protein release. Furthermore, the dynamic nanocomposite hydrogel exhibits good hemocompatibility and cytocompatibility. This work highlights the novel strategy for doping mussel inspired nanoparticles into dynamic hydrogel networks to enhance mechanical properties and sustain protein release.

BACKGROUND:Unexplained male infertility (UMI) accounts for 15%-30% of cases of male infertility, but it is poorly understood. We investigated potential risk factors for UMI leveraging data from a sperm bank with a "one male to multiple females" structure to reduce confounding arising from female factors. METHODS:Data on sperm donation provided by the Shanghai Human Sperm Bank (SHSB) to 39 qualified reproduction centres across China, along with information on IVF (In Vitro Fertilization) donor insemination cycles (DICs) performed at these centres and their reported outcomes, were retrieved from the Shanghai Human Sperm Bank (SHSB) for the period from 2004 to 2018. The association between semen parameters, demographic factors, and Cumulative Live Birth Rate from Donor Insemination (cLBR -DI) was analysed using linear regression analysis. Recurrent Failure of Sperm Donation IVF (RFDI) was defined as ≤1 pregnancy despite ≥4 DICs (indicating a cLBR -DI of ≤25%). Progressive Motility Rate (PR) refers to the percentage of grade (a + b) sperm among all sperm ((a + b)/(a + b + c + d)%). Forward Motility Rate (FR) refers to the percentage of grade (a + b) sperm among (a + b + c) sperm ((a + b)/(a + b + c)%). Sperm concentration<100 × 106/mL and FR > 95% was termed as normal group. The group with sperm concentration>200 × 106/mL or FR < 90% was termed high-risk RFDI (HR-RFDI). Logistic regression was used to assess correlations of RFDI. Propensity score matching (PSM) was used to match the RFDI and non-RFDI populations. Miscarriage conditions were also analysed in this study. FINDINGS:We included 4734 qualifying sperm donors and 17,307 IVF DICs, of which 2447 donors had more than 4 DICs per donor, forming a Pseudo-Polygyny Cohort. 8.05% of the 2447 donors with normal semen parameters and more than 4 DICs met RFDI criteria. RFDI donors exhibited higher sperm concentration (96.00 [75.00, 130.00] vs. 90.00 [72.00, 122.22], p = 0.057) and lower Forward Motility Rate (95.12 [89.02, 97.01] vs. 96.59 [93.33, 97.30], p < 0.001) compared to non-RFDI. Forward Motility Rate, rather than Progressive Motility Rate, correlated with cLBR -DI Compared to the normal group, HR-RFDI had an over three-fold higher risk of RFDI before (RR = 3.48, 95% CI [1.94, 6.25], p < 0.0001) and after (OR = 3.04, 95% CI [1.92, 4.81], p < 0.0001) PSM adjustment. Notably, RFDI donors had higher miscarriage rates (10.80 ± 14.20% vs. 3.90 ± 8.50%, p < 0.0001) compared to non-RFDI donors. High-risk RFDI donors also had higher miscarriage rates (5.40 ± 9.20% vs. 4.20 ± 9.40%, p = 0.011) compared to low-risk RFDI donors. INTERPRETATION:The negative impact of high sperm concentration on pseudo-polygyny cohort in vitro fertilization (IVF) outcomes strongly supports the hypothesis of a potential "inverted U-shape" relationship between sperm concentration and fertility. The superior correlation of FR with sperm donation IVF outcomes compared to PR suggests that FR may be a more meaningful measure of sperm motility in this context. The identification of an RFDI population, characterized by recurrent IVF failure and high miscarriage rates despite normal semen parameters, provides substantial evidence that the prevalence and severity of unexplained male infertility (UMI) may be greater than currently recognized. Therefore, a more comprehensive evaluation of male fertility should be considered in couples experiencing these issues. FUNDING:National Key R&D Program of China (2023YFC2705503), Fujian Province's Third Batch of Flexible Introduction of High-Level Medical Talent Teams (TD202307) and the National Key Technologies R&D Program (2023YFC2306700).

Purpose:Thyroid ultrasound is a primary tool for screening thyroid nodules (TNs), but existing risk stratification systems have limitations. Nowadays, machine learning (ML) offers advanced capabilities to handle high-dimensional data and complex patterns. This study aimed to develop an ML model integrating clinical data and ultrasound features to improve personalized prediction of TN malignancy. Methods:Data from 2,014 patients with TNs (2018.01-2024.01) were retrospectively analyzed, with 1,612 in the training set and 402 in the test set. Features included demographic, ultrasound, and thyroid function indices. Random Forest (RF) and Lasso regression were used for feature selection. Furthermore, six ML models (KNN, Logistic Regression, RF, Classification Tree, SVM, and XGBoost) were developed and validated via 10-fold cross-validation, evaluating performance using area under the receiver operating characteristic curve (AUC), accuracy, sensitivity, specificity, calibration curves, and decision curve analysis (DCA). Results:17 variables were influential factors for diagnosing TNs. All six models exhibited satisfactory predictive performance, with their accuracy ranging from 0.761 to 0.851 and AUC from 0.755 to 0.928. Among them, the XGBoost model demonstrated the best performance, achieving an AUC of 0.928, accuracy of 0.851, sensitivity of 0.933, and specificity of 0.650. Calibration curves showed strong agreement between predicted and observed malignancy probabilities, and DCA indicated net clinical benefit across a wide risk threshold range (0.2-0.9). Additionally, we have developed the model as a web-based calculator to facilitate its practical application. Conclusions:The XGBoost model effectively integrates multi-modal data to predict TN malignancy, offering improved accuracy and clinical utility.

Storage and transportation of dynamic hydrogel in ambient conditions is challenging. At present, cold-chain management is the most suitable option but it requires substantial infrastructure and energy. Moreover, the raw materials for preparing dynamic hydrogels often require chemical modification and purification, making the process cumbersome and time-consuming. This poses challenges for the practical applications of dynamic hydrogels. Here, we have developed a dynamic micron-sized (DMS) gel powder that can be packed, stored, and transported in a freeze-dried powdered form and can be rehydrated into a dynamic hydrogel. The dynamic hydrogel obtained by rehydrating the DMS gel powder stored under ambient conditions for 7 months can achieve properties similar to the original hydrogel, including stable protein delivery, mechanical properties, injectability, self-healing and remodeling. We propose a novel strategy for preparing a long-term storable DMS gel powder that can maintain properties similar to those of the original hydrogel. Considering the advantages in terms of convenience and commercialization for practical applications in the biomedical fields, this unique ready-to-use DMS gel powder offers a practical, scalable, and simple solution to enable mass production, cold-chain free, and low cost off-the-shelf dynamic hydrogel.

Customizable and viscoelastic porous biomaterials are highly desired as implant scaffold for repairing large-volume defects. Herein, we report customizable chitosan microfibers (CMFs)-based hydrospongels with mechanical properties comparable to soft tissues. The CMFs formed under high-speed shearing during crystallization of chitosan, and then they are crosslinked through covalent bond and hydrogen bond to form hydrospongels. The relatively high rigidity of CMFs provided hydrospongels with bulk elasticity at small deformation like hydrogel, and rapid reabsorb water in porous structures similar to sponge, endows excellent self-recovery property to hydrospongels after a large compression deformation. The optimized hydrospongel chemical crosslinked by glycerol triglycidyl ether (GTE) exhibits rapid recovery (1 s) owing to the synergistic effect of strong covalent and hydrogen bonds in porous structure. CMFs suspension has obvious shearing thinning property and can self-support its architecture after extrusion, realizing customizable capability to fabricate various 2D and 3D architectures. In vitro hemolysis test, cell test and CCK-8 assay demonstrate that hydrospongel has excellent hemocompatibility, cytocompatibility and non-toxicity. In vivo subcutaneous implantation of hydrospongel in rats reveal biodegradability and no inflammatory reaction. These properties provide a facile approach to fabricate green and mass-producible hydrospongel as an implant scaffold for biomedical applications.

In the period between surgery and systemic therapy for breast cancer, residual tumor cells may proliferate, leading to tumor recurrence. Additionally, intraoperative wound bleeding may cause surgical failure or the spread of tumor cells. This study introduces an innovative injectable hydrogel composed of oxidized hyaluronic acid (OHA) loaded 5-fluorouracil (5-FU) and N-carboxyethyl chitosan (CEC), designed for intraoperative hemostasis and tumor suppression in intraoperative breast cancer. The CEC/OHA injectable hydrogel was synthesized through a Schiff base reaction between the aldehyde group of OHA and the amino group of CEC, incorporating 5-FU during hydrogel formation. This CEC/OHA injectable hydrogel demonstrated hemostatic effects comparable to gelatin sponges in both an in vivo rat liver hemorrhage model and an in vitro rat tail amputation model. When loaded with 5-FU, the injectable hydrogel effectively inhibited the proliferation of MDA-MB-231 breast cancer cells in vitro, significantly inhibited tumor growth and recurrence in vivo, and did not induce significant damage or inflammatory response in any major organ. This CEC/OHA & 5-FU injectable hydrogel is envisioned as a complementary therapeutic regimen during the intraoperative period in breast cancer surgery to prevent hemostasis and tumor recurrence.

Red fluorescent hydrogels possessing injectable and self-healing properties have widespread potential in biomedical field. It is still a challenge to achieve a biomacromolecules based dynamic hydrogels simultaneously combining with excellent red fluorescence, good mechanical properties, and biocompatibility. Here we firstly exploited hydrophilic inclusion complex of (R-CDs@α-CD) derived from red fluorescent carbon dots (R-CDs) and α-cyclodextrin (α-CD), and then achieved a red fluorescent and dynamic polysaccharide R-CDs@α-CD/CEC-l-OSA hydrogel. The nanocomposite hydrogel can be fabricated through controlling dopant of red fluorescent R-CDs@α-CD into dynamic polymer networks, taking dynamic crosslinked N-carboxyethyl chitosan (CEC) and oxidized sodium alginate (OSA) as an example. The versatile hydrogel simultaneously combines the features of injection, biocompatibility, and augmented mechanical properties and self-healing behavior, especially in rapid self-recovery even after integration. The R-CDs@α-CD uniformly dispersed into dynamic hydrogel played the role of killing two birds with one stone, that is, endowing red fluorescence as a hydrophilic fluorescent substance, and improving mechanical and self-healing properties as a dynamic nano-crosslinker, via providing hydrogen bonds as reversible crosslinkings. The novel red fluorescent and dynamic hydrogel based on polysaccharides hold promise for using as biomaterials in biomedical field.

The fabrication of scaffolds capable of the sustained release of the vascular endothelial growth factor (VEGF) to promote angiogenesis for a long time remains a challenge in tissue engineering. Here, we report a facile approach for effectively fabricating a bioactive scaffold that gradually releases VEGF to promote angiogenesis. The scaffold was fabricated by coating polydopamine (PDA) on a konjac glucomannan (KGM) scaffold, followed by the surface immobilization of VEGF with PDA. The resulting VEGF-PDA/KGM scaffold, with a porous and interconnected microstructure (392 mu m pore size with 84.80 porosity), combined the features of long-term biodegradability (10 weeks with 51 % degradation rate), excellent biocompatibility, and sustained VEGF release for up to 21 days. The bioactive VEGF-PDA/KGM scaffold exhibited multiple angiogenic activities over time, as confirmed by in vivo and in vitro experiments. For example, the scaffold significantly promoted the attachment and proliferation of human umbilical vein endothelial cells and the formation of vascular tubes in vitro . Moreover, the in vivo results demonstrated the formation and maturation of blood vessels after subcutaneous implantation in rats for four weeks. This promising strategy is a feasible approach for producing bioactive materials that can induce angiogenesis in vivo . These findings provide a new avenue for designing and fabricating biocompatible and long-term biodegradable scaffolds for sustained VEGF release to facilitate angiogenesis.

Objective: Metaplastic breast carcinoma (MBC) is a special type of morphologically heterogeneous and aggressively invasive breast cancer. MBC is characterized by the transformation of tumor epithelium into squamous epithelium and/or mesenchymal components, including differentiation into spindle cells, chondrocytes, and osteocytes. Due to its rarity and invasiveness, there is a paucity of research on MBC prognosis. Furthermore, there are currently no treatment guidelines for MBC. This study analyzed the clinicopathological characteristics, immunophenotype, and prognostic features of MBC. Our aim was to better characterize MBC, thereby identifying potential prognostic factors and new treatment methods. Moreover, we also describe an MBC case treated experimentally with anti-vascular targeted therapy. Material and Methods: We retrospectively analyzed clinical pathological data on 54 female patients with MBC from Shaanxi Provincial People's Hospital and the XiJing Hospital of Air Force Medical University. These cases were diagnosed with MBC between January 1(st), 2013, and October 1(st), 2018. All patients were from the northwest region of China. The gross morphological, histological, and immunohistochemical features of MBC were analyzed. Kaplan-Meier analysis was used to calculate the survival rate, and univariate analysis was performed to identify significant prognostic factors. In addition, the treatment of an MBC patient with anti-angiogenic therapy was described, and a relevant literature review was conducted. Results: MBC was diagnosed in 32 left breasts and 22 right breasts from 54 women aged 21-76 years (median age of 57 years). The maximum tumor diameter ranged from 0.6 to 14 cm (average of 4.1 cm). Of the 54 patients, 47 underwent surgical treatment, with lymph node metastasis found in 17.0% (8/47). According to the World Health Organization classification criteria for breast tumors, the study cohort consisted of 15 cases of squamous cell carcinoma, ten cases of spindle cell carcinoma, nine cases of carcinoma with associated stromal differentiation, 18 cases of mixed carcinoma, and two cases of adenocarcinoma with squamous differentiation. Based on the American Joint Committee on Cancer clinical staging criteria, the patients were classified as Stage I (10 cases, 18.5%), Stage II (26 cases, 48.1%), Stage III (11 cases, 20.4%), and Stage IV (7 cases, 13.0%). Immunohistochemical analysis revealed that 94.4% of patients had triple-negative breast cancer (TNBC), 47 cases showed mutant tumor protein 53 (TP53) expression, 29 cases showed positive epidermal growth factor receptor (EGFR) expression, 43 cases showed positive E-cadherin expression, and 37 cases showed positive Cluster of Differentiation 24 expression. The Ki-67 index ranged from 20% to 90%. Univariate analysis showed that the Ki-67 index was not significantly associated with either progression-free survival (PFS) or overall survival (OS) in MBC patients. Patients with negative axillary lymph nodes had significantly better PFS and OS than those with positive nodes (P < 0.05), and patients with clinical stage I-II disease had better PFS and OS than those with stage III-IV disease (P < 0.05). Patients treated with anthracycline-containing chemotherapy had significantly better PFS than those who did not receive chemotherapy. Univariate analysis revealed that the high expression of EGFR correlated with worse PFS (P < 0.05). The type of surgical approach employed did not affect the prognosis of MBC patients. Following the application of anti-angiogenic therapy, a rapid partial response was observed in an MBC patient with carcinoma and associated stromal differentiation. This patient subsequently underwent surgery and radiation therapy and has now achieved over 6 years of PFS. Conclusion: MBC is a heterogeneous group of tumors with high malignancy and poor prognosis. The large majority is TNBC and exhibits unique immune phenotypes. The poor PFS of MBC patients may be related to EGFR expression, which could become a potential therapeutic target in these patients. Surgery remains the primary treatment method for MBC. The present study found that sentinel lymph node biopsy was feasible in appropriate patients, and that chemotherapy regimens incorporating anthracycline-class drugs did not appear to improve OS. Anti-angiogenic therapy holds promise as a potentially effective treatment approach for MBC, and the optimization of systemic treatment strategies should be a priority in the management of these patients.

This study aimed to develop prognostic prediction models for patients diagnosed with synchronous thyroid and breast cancer (TBC). Utilizing the SEER database, key predictive factors were identified, including T stage of thyroid cancer, T stage of breast cancer, M stage of breast cancer, patient age, thyroid cancer surgery type, and isotope therapy. A nomogram predicting 5-year and 10-year survival rates was constructed and validated, exhibiting strong performance (C-statistic: 0.79 in the development cohort (95% CI: 0.74-0.84), and 0.82 in the validation cohort (95% CI: 0.77-0.89)). The area under the Receiver Operator Characteristic (ROC) curve ranged from 0.798 to 0.883 for both cohorts. Calibration and decision curve analyses further affirmed the model's clinical utility. Stratifying patients into high-risk and low-risk groups using the nomogram revealed significant differences in survival rates (P < 0.0001). The successful development and validation of this nomogram for predicting 5-year and 10-year survival rates in patients with synchronous TBC hold promise for similar patient populations, contributing significantly to cancer research.

Dynamic hydrogels with the features of injection, self-healing, and remodeling at the target site have been developed as smart multifunctional biomaterials for drug delivery. However, most self-healing injectable hydrogels are difficult to control protein release after implantation, owing to the deficiency of pH responsiveness, which reduces the bioavailability of proteins. Herein, we propose a facile strategy to endow pH responsiveness into a dynamic hydrogel with both self-healing and injectable capabilities, by crosslinking biomacromolecular backbones via dual pH sensitive dynamic covalent bond. Particularly, oxidized konjac glucomannan (OKGM) can be crosslinked with poly (aspartic hydrazide) (PAHy) and N-carboxyethyl chitosan (CEC) to form dynamic acylhydrazone bonds and imide bonds, respectively, endowing the hydrogel with pH responsiveness and dynamic behaviors. Specifically, PAHy facilitates the formation of acylhydrazone bonds, improving the mechanical properties and pH sensitivity while reducing the degradation behavior of the hydrogels under physiological conditions. Kinetics indicate that the release of bovine serum albumin follows Fick diffusion under different pH conditions. The pH responsive hydrogel with self-healing injectable capabilities has the potential to be used as a controllable and sustain release carrier for protein drugs.

OBJECTIVES:The objective of this study was to develop and validate a nomogram model integrating clinical, biochemical and ultrasound features to predict the malignancy rates of Thyroid Imaging Reporting and Data System 4 (TR4) thyroid nodules. METHODS:A total of 1557 cases with confirmed pathological diagnoses via fine-needle aspiration (FNA) were retrospectively included. Univariate and multivariate logistic regression analyses were conducted to identify independent predictors of malignancy. These predictors were incorporated into the nomogram model, and its predictive performance was evaluated using receiver-operating characteristic curve (AUC), calibration plots, net reclassification improvement (NRI), integrated discrimination improvement (IDI) and decision curve analysis (DCA). RESULTS:Eight out of 22 variables-age, margin, extrathyroidal extension, halo, calcification, suspicious lymph node metastasis, aspect ratio and thyroid peroxidase antibody-were identified as independent predictors of malignancy. The calibration curve demonstrated excellent performance, and DCA indicated favourable clinical utility. Additionally, our nomogram exhibited superior predictive ability compared to the current American College of Radiology (ACR) score model, as indicated by higher AUC, NRI, IDI, negative likelihood ratio (NLR) and positive likelihood ratio (PLR) values. CONCLUSIONS:The developed nomogram model effectively predicts the malignancy rate of TR4 thyroid nodules, demonstrating promising clinical applicability.

Abstract Background Mycobacteria bloodstream infections are common in immunocompromised people and usually have disastrous consequences. As the primary phagocytes in the bloodstream, monocytes and neutrophils play critical roles in the fight against bloodstream mycobacteria infections. In contrast to macrophages, the responses of monocytes infected with the mycobacteria have been less investigated. Results In this study, we first established a protocol for infection of non-adherent monocyte-like THP-1 cells (i.e. without the differentiation induced by phorbol 12-myristate 13-acetate (PMA) by bacillus Calmette-Guérin (BCG). Via the protocol, we were then capable of exploring the global transcriptomic profiles of non-adherent THP-1 cells infected with BCG, and found that NF-κB, MAPK and PI3K-Akt signaling pathways were enhanced, as well as some inflammatory chemokine/cytokine genes (e.g. CCL4, CXCL10, TNF and IL-1β) were up-regulated. Surprisingly, the Akt-HIF-mTOR signaling pathway was also activated, which induces trained immunity. In this in vitro infection model, increased cytokine responses to lipopolysaccharides (LPS) restimulation, higher cell viability, and decreased Candida albicans loads were observed. Conclusions We have first characterized the transcriptomic profiles of BCG-infected non-adherent THP-1 cells, and first developed a trained immunity in vitro model of the cells.

Trained immunity is one of the mechanisms by which BCG vaccination confers persistent nonspecific protection against diverse diseases. Genomic differences between the different BCG vaccine strains that are in global use could result in variable protection against tuberculosis and therapeutic effects on bladder cancer. In this study, we found that four representative BCG strains (BCG-Russia, BCG-Sweden, BCG-China, and BCG-Pasteur) covering all four genetic clusters differed in their ability to induce trained immunity and nonspecific protection. The trained immunity induced by BCG was associated with the Akt-mTOR-HIF1α axis, glycolysis, and NOD-like receptor signaling pathway. Multi-omics analysis (epigenomics, transcriptomics, and metabolomics) showed that linoleic acid metabolism was correlated with the trained immunity–inducing capacity of different BCG strains. Linoleic acid participated in the induction of trained immunity and could act as adjuvants to enhance BCG-induced trained immunity, revealing a trained immunity–inducing signaling pathway that could be used in the adjuvant development.

The Klebsiella pneumoniae (K. pneumoniae, Kp) populations carrying both resistance-encoding and virulence-encoding mobile genetic elements (MGEs) significantly threaten global health. In this study, we identified a new anti-CRISPR gene (acrIE10) on a conjugative plasmid with self-target sequence in K. pneumoniae with type I-E* CRISPR-Cas system. AcrIE10 interacts with the Cas7* subunit of K. pneumoniae I-E* CRISPR-Cas system. The crystal structure of the AcrIE10-KpCas7* complex suggests that AcrIE10 suppresses the I-E* CRISPR-Cas by binding directly to Cas7 to prevent its hexamerization, thereby preventing the surveillance complex assembly and crRNA loading. Bioinformatic and functional analyses revealed that AcrIE10 is functionally widespread across diverse species. Our study reports a novel anti-CRISPR and highlights its potential role in spreading resistance and virulence among pathogens.

Biomacromolecules based injectable and self-healing hydrogels possessing high mechanical properties have widespread potential in biomedical field. However, dynamic features are usually inversely proportional to toughness. It is challenging to simultaneously endow these properties to the dynamic hydrogels. Here, we fabricated an injectable nanocomposite hydrogel (CS-NPs@OSA-l-Gtn) stimultaneously possessing excellent autonomous self-healing performance and high mechanical strength by doping chitosan nanoparticles (CS-NPs) into dynamic polymer networks of oxidized sodium alginate (OSA) and gelatin (Gtn) in the presence of borax. The synergistic effect of the multiple reversible interactions combining dynamic covalent bonds (i.e., imine bond and borate ester bond) and noncovalent interactions (i.e., electrostatic interaction and hydrogen bond) provide effective energy dissipation to endure high fatigue resistance and cyclic loading. The dynamic hydrogel exhibited excellent mechanical properties like maximum 2.43 MPa compressive strength, 493.91 % fracture strain, and 89.54 kJ/m3 toughness. Moreover, the integrated hydrogel after injection and self-healing could withstand 150 successive compressive cycles. Besides, the bovine serum albumin embedded in CS-NPs could be sustainably released from the nanocomposite hydrogel for 12 days. This study proposes a novel strategy to synthesize an injectable and self-healing hydrogel combined with excellent mechanical properties for designing high-strength natural carriers with sustained protein delivery.

Anti-HER2 therapy has significantly improved the survival rates of patients with HER2+ breast cancer. However, a subset of these patients eventually experience treatment failure, and the underlying genetic mechanisms remain largely unexplored. This underscores the need to investigate the genomic heterogeneity of HER2+ breast cancer. In this study, we focus on HER2+/HR- breast cancer, as it differs from HER2+/HR+ breast cancer in terms of genetic and biological characteristics. We performed gene-targeted genome sequencing on 45 HER2+/HR- breast cancer samples and identified 650 mutations across 268 cancer-related genes. TP53 (71.1%) and PIK3CA (35.6%) were the most frequently mutated genes in our sample. Additionally, ERBB2 (77.8%), CDK12 (42.2%), and MYC (11.1%) exhibited a high frequency of copy number amplifications (CNAs). Comparative analysis with two other HER2+/HR- breast cancer cohorts revealed that our cohort had higher genetic variation rates in ARID1A, PKHD1, PTPN13, FANCA, SETD2, BRCA2, BLM, STAG2, FAT1, TOP2A, POLE, ATM, KMT2B, FGFR4, and EPAS1. Notably, in our cohort, NF1 and ATM mutations were more prevalent in trastuzumab-resistant patients (NF1, p=0.016; ATM, p=0.006) and were associated with primary trastuzumab resistance (NF1, p=0.042; ATM, p=0.021). Moreover, patients with NF1 mutations (p=0.009) and high histological grades (p=0.028) were more likely to experience early relapse. Ultimately, we identified a unique cancer-related gene mutation profile and a subset of genes associated with primary resistance to trastuzumab and RFS in patients with HER2+/HR- breast cancer in Northwest China. These findings could lay the groundwork for future studies aimed at elucidating the mechanisms of resistance to trastuzumab and improving HER2-targeted treatment strategies.