Perfluorooctanoic acid (PFOA) resists conventional oxidation due to its strong C-F bonds. We hypothesized that sulfurization might improve zero-valent iron’s ability to activate peroxymonosulfate (PMS) for PFOA degradation. Here we show that a hydrothermally synthesized sulfurized nanoscale zero-valent iron (S-nZVI, S: Fe = 1: 10) coupled with PMS degraded PFOA (1 mg/L) and released fluoride during a 360 min reaction. The S-nZVI/PMS system substantially outperformed PMS or S-nZVI alone. After 360 min, the residual PFOA fraction fell to ~ 0.15, and fluoride release reached 47.33% (17.1 µM F−, 0.326 mg/L) - evidence of C-F bond cleavage. Higher PMS dosage improved degradation; the optimal S-nZVI loading was near 0.5 g/L. Acidic conditions favored the reaction. Mechanistic experiments (quenching, oxygen-control, ROS analysis, PMS consumption and EPR) indicated a radical/non-radical hybrid process involving •OH/SO4•−-type radical reactions, O2•−-related chemistry and a 1O2-mediated non-radical pathway. Shorter-chain intermediates (PFHpA, PFHxA, PFPeA and PFBA) confirmed stepwise chain shortening, while fluorine mass balance showed that fluoride, residual PFOA and detected intermediates accounted for most of the initial fluorine. Real waters (tap and surface water) inhibited the reaction due to dissolved organic matter and inorganic ions, yet the system still removed about half of the PFOA. S-nZVI remained active over multiple cycles and also degraded GenX, albeit more slowly than PFOA. These results identify sulfurized nZVI as a promising PMS activator for PFAS treatment under environmentally relevant conditions.

Autoimmune encephalitis (AE) comprises clinically and immunopathologically heterogeneous central nervous system disorders whose early recognition should be syndrome-based and should not depend on antibody results alone. Cerebellar and brainstem manifestations occur in selected AE and related central nervous system neuroimmune disorders, but their frequency, anatomical specificity, and mechanisms remain incompletely defined. In this narrative Review, we use the cerebellum-brainstem network as an anatomical and clinical organizing framework rather than proposing a discrete anatomical axis or a new disease entity. In selected neuronal-surface-antibody disorders, antibodies can directly alter receptor trafficking, receptor availability, protein interactions, or synaptic transmission, whereas intracellular-antigen-associated and paraneoplastic syndromes are more often linked to cytotoxic T-cell-dominant neuronal injury. To stabilize disease scope, central AE and directly relevant central nervous system autoimmune syndromes form the core evidence base; immune-mediated cerebellar ataxias, paraneoplastic syndromes, acute disseminated encephalomyelitis, and myelin oligodendrocyte glycoprotein antibody-associated disease are included only when they provide direct infratentorial evidence; and Miller Fisher syndrome and Guillain-Barré syndrome are retained solely as peripheral anatomical comparators, whereas Bickerstaff brainstem encephalitis represents a central brainstem syndrome. We present a hypothesis-generating circuit framework linking immune target engagement to cerebellar output and connected brainstem manifestations, while explicitly marking extrapolations from non-AE models as hypotheses [H]. We also distinguish a predominantly functional pattern from an established structural-injury pattern as non-sequential research constructs rather than stages, biomarker-defined transitions, treatment windows, or clinical algorithms. Current imaging studies demonstrate that infratentorial metabolic and structural abnormalities can occur, but no reproducible cerebellum-brainstem diagnostic, prognostic, or treatment-selection signature has been prospectively validated.

Mesenchymal stem cells (MSCs) are widely used in cell therapies; however, replicative senescence during in vitro expansion limits their potency and consistency. Partial reprogramming has emerged as a promising strategy to rejuvenate aged cells, yet concerns regarding unintended pluripotent conversion have hindered its translational application. Here we developed a circular RNA (circRNA)-based platform enabling safe partial reprogramming and rejuvenation of MSCs. We generated a lipid nanoparticle (LNP)-delivered circRNA co-expressing OCT4, SOX2, and KLF4 (circ-OSK) to achieve transient and coordinated expression of partial reprogramming factors without genomic modification in MSCs. Using a rigorously validated droplet digital PCR (ddPCR) system targeting the pluripotency marker ESRG, we demonstrated that circ-OSK delivery alone is insufficient to transform MSCs into induced pluripotent stem cells (iPSCs). Circ-OSK-mediated partial reprogramming significantly rejuvenated senescent late-passage MSCs by reducing senescence-associated β-galactosidase activity, DNA damage markers, and inflammatory cytokine secretion, while preserving canonical MSC surface markers and genome stability, and restoring proliferative, migratory and tri-lineage differentiation capacities at a level comparable to early-passage MSCs. The rejuvenated MSCs generated through this process are defined as partially reprogrammed MSCs (prMSCs). Further RNA-seq analysis revealed that prMSCs restored youthful gene expression and promoted a stress resilient and regenerative transcriptional program via repressing the PI3K-Akt signaling pathway. Combined RNA-seq and SNP array analysis confirmed circ-OSK-mediated partial reprogramming retained MSC identity and genome integrity without pluripotent conversion. Our study has established a circRNA-mediated partial reprogramming strategy that rejuvenates MSCs without pluripotent conversion, offering a safety-oriented approach to enhance the potency and quality of MSC-based cell therapy products.

The mucosal system, which includes the respiratory, gastrointestinal, and urogenital tracts, serves as a primary entry point for pathogens, with a unique immune microenvironment and specialized defense mechanisms. In recent years, especially following the onset of the COVID-19 pandemic, there has been increasing recognition of the importance of mucosal immunity, motivated by an enhanced comprehension of its fundamental mechanisms. Currently, strategies based on mucosal delivery systems to administer antigens and induce strong mucosal protective immunity have become a key focus in the development of mucosal vaccines. Compared with conventional intramuscular delivery, mucosal vaccination can simultaneously elicit a robust local mucosal response, effectively block pathogen entry into the local mucosa, and generate systemic immune responses to prevent symptomatic infections and severe disease. In addition, mucosal delivery offers advantages such as ease of administration and low invasiveness, making it a more widely acceptable approach to vaccination. In the present study, we conducted a systematic review of the mechanisms of mucosal immunity, the technological platforms for mucosal vaccines, and proposed a perspective on the challenges and future directions for the development of next-generation mucosal vaccines, with the goal of enhancing public knowledge and awareness regarding mucosal immunity and its possible effects on global health.

Human granulocyte colony-stimulating factor (hG-CSF) is primarily used to treat neutropenia induced by cancer chemotherapy and bone marrow transplantation. The current identification test for hG-CSF relies on Western blot (WB), a labor-intensive and technically demanding method. This study aimed to screen and prepare an anti-hG-CSF nanobody to identify and quantify hG-CSF, with the ultimate goal of developing colloidal gold-labeled nanobody test strips for rapid identification. An alpaca was immunized with hG-CSF, and the VHH gene sequence encoding the anti-hG-CSF nanobody was obtained through sequencing following phage display library construction and multiple rounds of biopanning. The nanobody C68, obtained from screening, was expressed by E. coli, and its physicochemical properties such as molecular weight, isoelectric point, and affinity were characterized after purification. WB analysis demonstrated excellent performance of the nanobody in identification tests in terms of specificity, limit of detection (LOD), applicability with products from various manufacturers, and thermal stability. Additionally, we established an ELISA method for hG-CSF quantification utilizing the nanobody C68 and conducted methodological validation. Finally, colloidal gold-based test strips were constructed using the nanobody C68, with a LOD of 30 μg/mL, achieving rapid identification for hG-CSF. This study represents a novel application of nanobodies in pharmaceutical testing and offers valuable insights for developing identification tests for other recombinant protein drugs.

Glucagonoma, a rare neuroendocrine tumor, lacks targeted treatment drugs. Excessive secretion of glucagon is the main cause of its clinical syndrome. To explore targeted therapeutic drugs that can inhibit glucagon secretion and tumor proliferation, we investigated the effect of Trametenolic Acid (TA) on mouse pancreatic alpha TC1 clone 6 (αTC1-6) cells and its regulatory role in the PI3K/AKT signaling pathway. Cell viability of αTC1-6 cells was assessed via the MTT assay. Glucagon content in cell culture supernatants was measured using an Enzyme-Linked Immunosorbent Assay (ELISA). Autophagic vacuoles were visualized through Monodansylcadaverine (MDC) staining. The expression of autophagy-related proteins including Atg7, LC3 Ⅱ and PI3K/AKT signaling pathway-related proteins mTOR and FoxO1 were determined by Western blot. The results showed that the proliferation of αTC1-6 cells was significantly inhibited by TA in a dose- and time-dependent manner, and the IC50 was 140.71, 26.77 and 1.99 μM after treatment of 12, 24, and 48 h, respectively. The secretion of glucagon was significantly inhibited by TA. The MDC staining results showed that the fluorescent labeled autophagic vesicles in the TA group were increased. The Western blot results showed that the expression of Atg7 and LC3 Ⅱ was promoted by TA in a dose-dependent manner, the phosphorylation of PI3K, AKT, mTOR and FoxO1 was significantly inhibited, and the expression of FoxO1 protein was increased. These results demonstrated that TA can inhibit glucagon secretion, induce autophagy, and suppress cell proliferation in αTC1-6 cells. The mechanism may be associated with the PI3K/AKT signaling pathway.

Recently, rapidly evolving STING-based immunotherapies have offered novel therapeutic options for various cancer types. However, systemic administration of STING agonists raises safety concerns, and intratumoral injection is constrained by tumor accessibility. Herein we developed an immune-stimulating antibody conjugate (ISAC) that links STING agonists to antibodies that target HER2-positive tumor cells via a cleavable linker. In vivo studies demonstrated that the STING agonist ISAC is well tolerated and exhibits potent antitumor activity in syngeneic mouse tumor models. Investigations in STING-knockout HER2-positive tumor cells and STING-knockout mouse models revealed that the STING pathway primarily mediates antitumor effects upon the activation of immune and tumor cells and that the activation of immune cells plays a stronger role. Additionally, our findings indicate that the STING agonist ISAC enhances both innate and adaptive antitumor immune responses, leading to sustained antitumor activity and the establishment of immune memory. These outcomes support the clinical development of the STING agonist ISACs.

Therapeutic cancer vaccines are a new growth point of biomedicine with broad industrial prospects in the post-COVID-19 era. Many large international pharmaceutical companies and emerging biotechnology companies are deploying different tumor therapeutic cancer vaccine projects, focusing on promoting their clinical transformation, and the vaccine industry has strong momentum for development. Such vaccines are also the core engine and pilot site for the development of new vaccine targets, new vectors, new adjuvants, and new technologies, which play a key role in promoting the innovation and development of vaccines. Various therapeutic cancer vaccines, such as viral vector vaccines, bacterial vector vaccines, cell vector vaccines, peptide vaccines, and nucleic acid vaccines, have all been applied in clinical research. With the continuous development of technology, therapeutic cancer vaccines are evolving towards the trends of precise antigens, efficient carriers, diversified adjuvants, and combined applications. For instance, the rapidly advancing mRNA-4157 vaccine is a typical representative that combines personalized antigens with efficient delivery vectors (lipid nanoparticles, LNPs), and it also shows synergistic advantages in melanoma patients treated in combination with immune checkpoint inhibitors. In this article, we will systematically discuss the current research and development status and clinical research progress of various therapeutic cancer vaccines.

Pharmacokinetic study of SHR-2005 in B-hCD40 mice.

Thymic stromal lymphopoietin (TSLP) promotes Th2-mediated inflammation via dendritic cell activation and STAT5 signaling and plays critical roles in inflammatory disorders and allergic diseases. While tezepelumab stands as the first and only approved anti-TSLP monoclonal antibody (mAb) with over 10 anti-TSLP mAbs in clinical development, no validated anti-TSLP mAb bioassay was reported yet. Bioactivity determination is essential for ensuring mAb quality. To bridge this gap, we generated a novel HuT78-STAT5-luc reporter cell line through lentiviral transduction of STAT5 response element-driven luciferase into HuT78 cells. This stable cell line expressed luciferase in a TSLP dose-responsive manner. After systematic optimization of cell density, incubation time, TSLP concentration and mAb concentration, we established a reporter gene assay (RGA) with four-parameter regression compliance for the anti-TSLP mAb bioactivity determination. The RGA underwent full validation according to the International Council for Harmonization (ICH) Q2(R2) guideline, namely specificity, linearity, accuracy, precision and robustness. In conclusion, we established a robust and user-friendly RGA that can be applied for the quality control of anti-TSLP mAbs.

CD40 expression in normal human and rhesus tissues by immunohistochemistry.

The binding ability of 9E5 to TNF family members.

The ADCC and CDC activity of SHR-2005.

Effects of SHR-2005 on the release of various cytokines in human peripheral blood mononuclear cells (PBMC).

Oncolytic virus (OV) immunotherapy, particularly with oncolytic herpes simplex virus (oHSV), has become a promising new strategy in cancer treatment. This field has achieved significant clinical milestones, highlighted by the FDA approval of Talimogene laherparepvec (T-VEC) for melanoma in 2015 and the approval of Teserpaturev/G47Δ for malignant glioma in Japan in 2021. This review synthesizes the key preclinical and clinical advancements in oHSV therapy over the last decade, critically analyzing the core challenges in target selection, genetic modification, administration routes, and targeted delivery. Key findings indicate that arming oHSV with immunomodulatory transgenes, such as cytokines and antibodies, and combining it with immune checkpoint inhibitors are critical strategies for enhancing therapeutic efficacy. Future research will focus on precision engineering using CRISPR/Cas9, the development of novel delivery vehicles like nanoparticles and mesenchymal stem cells (MSCs), and biomarker-guided personalized medicine, aiming to provide safer and more effective solutions for refractory cancers. This review synthesizes oHSV advances and analyzes novel delivery and gene-editing strategies.

The activation of DC, B and T cells of SHR-2005 and APX005M analogue.

Binding of SHR-2005 to human FcgR as determined by SPR assay.

Mucosal immunity is crucial for preventing the infection and transmission of respiratory viruses. Nasal antibody is inversely correlated with a lower risk of infection with respiratory viruses. However, the current reference standard for nasal antibody assessment is serum-based, mainly consisting of monomeric IgG and IgA. The applicability of serum-derived standards for assessing nasal antibodies, consisting mostly of dimeric or polymeric secretory IgA (sIgA), remains unvalidated. Herein, we first proved that the sera-derived standard was not applicable for assessing nasal antibodies. Using a non-homologous standard as a calibrator introduced systematic error up to 10 times, which did not benefit the understanding of mucosal antibody response. Therefore, we attempted to develop two candidate standards (CS1, CS2) using nasal mucosal lining fluids (NMLFs) collected from SARS-CoV-2 Omicron convalescents or intranasal vaccine recipients, and CS3 using a sIgA monoclonal antibody. CS2 exhibited broad-spectrum binding activity against 12 SARS-CoV-2 strains, including all tested Omicron subvariants. A collaborative study conducted by seven laboratories demonstrated that CS2 improved the harmonization of inter-laboratory variability (pre-standardization geometric coefficients of variance, 14-314%; post-standardization, 3-35%). Using CS2 ensured an accurate assessment of nasal antibodies. Thus, CS2 was established as a national standard for evaluating nasal SARS-CoV-2-specific antibodies (Lot: 300052-202401, 1000 U/mL). Our work provides a benchmark for evaluating mucosal vaccines for SARS-CoV-2 and inspires new avenues for developing new reference standards for other mucosal vaccines.

The difference in the binding affinity of SHR-2005 to human and mouse FcgRIIb.

Gastrodia elata Blume (GEB) is a potential medicinal and edible plant with several active components and pharmacological activity that has a high application value in medicine and the food business. However, in natural conditions, GEB seed has a very low germination rate and depends on two specific fungi, germinal and nutritive fungi, to complete the germination process and growth. Armillaria mellea, while acting as a nutrient supplier, actually inhibits the germination of GEB seeds. Mycena strains, as the main germinating fungi, can facilitate germination but cannot support the subsequent growth and development of GEB. It requires symbiotic interactions with Mycena and Armillaria mellea to obtain nutrients for its complex life cycle. Our previous studies have shown that trametenolic acid (TA) can effectively promote seed germination of GEB. The aim of this study was to use transcriptome sequencing to further understand the potential mechanism of seed germination triggered by TA in GEB, in order to lay the groundwork for developing a new germination-growth system for GEB with Armillaria mellea. The untreated symbiotic group (Group A0) did not germinate in the seed germination test. The high-dose TA-treated symbiotic group (Group B), the low-dose TA-treated symbiotic group (Group C), and the non-symbiotic untreated germination group (Group A) had germination rates of 85.01, 61.18 and 27.39%, respectively. This indicates that TA treatment can induce symbiosis with Armillaria mellea in GEB seeds and significantly increase germination rates. Transcriptome sequencing (RNA-seq) of Groups A, B, and C identified 86,843 annotated genes. There were more down-regulated genes than up-regulated genes, with 3912, 2518, and 814 differentially expressed genes (DEGs) between B and A, C and A, and B and C, respectively. The DEGs were mainly involved in DNA transcription factors, cell wall actions, plant-pathogen interactions, phenylpropanoid biosynthesis, phytohormone signal transduction, and starch-sucrose metabolism pathways. Six genes were confirmed using qRT-PCR: Down-regulated genes in the lignin biosynthesis pathway include MYB4 and 4CL, while GA20ox1 in the gibberellin biosynthesis pathway was also down-regulated. Up-regulated genes in the plant-pathogen interaction pathway are AIB and WRKY51, with MYB44 in the lignin biosynthesis pathway showing up-regulation. The transcriptomics results supported these expression patterns. Lignin, GA, and abscisic acid (ABA) levels were analyzed in GEB protocorms to understand how TA promotes germination. Results showed that groups B and C had lower lignin and ABA levels, but higher GA levels compared to group A. The study revealed that certain genes play a crucial role in promoting GEB seed germination through TA, by regulating gene expression to alter lignin content and hormone levels, breaking seed dormancy, facilitating seed-fungus interactions, and promoting symbiotic relationships with Armillaria mellea. TA modulates the expression of genes involved in lignin biosynthesis and hormone signaling, leading to an increase in GA content and a decrease in ABA and lignin content. This helps seeds break dormancy and promote germination. Additionally, TA can enhance GEB's defense response against fungi by regulating plant-pathogen interaction genes. It also improves the interactions between GEB and Armillaria mellea, overcoming the technical challenges associated with using Armillaria mellea as a germinating fungus. This establishes a new symbiotic germination-growth system between Armillaria mellea and GEB, laying the foundation for further research on the molecular mechanisms of GEB seed germination.