Huricha Baigude

Background: Costunolide (Cos) is a sesquiterpene lactone isolated from Saussurea lappa with reported anti-inflammatory properties. Its direct molecular targets in neuroinflammatory responses, however, have not been fully defined. Methods: LPS-stimulated BV2 microglia were used to examine the effect of Cos on neuroinflammatory activation. Candidate binding proteins were screened using MetPro and further assessed by molecular docking. The roles of selected targets were tested by siRNA knockdown, while direct binding was examined using surface plasmon resonance (SPR). Transcriptome sequencing and qPCR were used to evaluate downstream inflammatory gene changes. Results: Cos reduced LPS-induced phosphorylation of NF-κB p65, with little change in total p65 protein levels. MetPro analysis pointed to G3BP1 and APRT as potential Cos-associated proteins, and docking analysis supported possible binding of Cos to both targets. Knockdown of either G3BP1 or APRT reduced the inhibitory effect of Cos on p65 phosphorylation. SPR assays confirmed direct interaction between Cos and G3BP1/APRT. Transcriptomic analysis, together with qPCR validation, showed that Cos affected LPS-responsive inflammatory transcriptional programs. Conclusion: Cos suppresses microglial inflammatory activation partly through G3BP1 and APRT, with downstream inhibition of NF-κB-related signaling. These results provide mechanistic evidence for the anti-neuroinflammatory activity of Cos and support further investigation of its therapeutic potential.

Cationic curdlan is a promising siRNA delivery platform because it protects siRNA, promotes intracellular siRNA release through charge reduction, prolongs intracellular retention, and supports sustained gene-silencing. However, its relatively high cationic charge can induce cytotoxicity and limit its therapeutic potential. In this study, to reduce toxicity while enhancing siRNA delivery efficiency, we conjugated short ethylene glycol side chains to 6-amino-6-deoxy curdlan and compared the siRNA transfection efficiency of the resulting polymers with that of the parental polymer. Conjugation of ethylene glycol side chains significantly improved both the biocompatibility and in vitro siRNA delivery efficiency, yielding polymers designated MEG1 and MEG2. Notably, conjugation of a short ethylene glycol side chain terminated with a lysine residue (designated CegK1) resulted in markedly enhanced biocompatibility and siRNA delivery efficiency in multiple human glioblastoma cell lines, including U251, U87-MG, and T98G. These findings demonstrate that short ethylene glycol side-chain modification is an effective strategy to optimize cationic curdlan as a safe and efficient siRNA delivery carrier for glioblastoma therapy.

Background: Aconitum kusnezoffii Reichb. is a commonly used medicinal herb in Mongolian medicine (a branch of traditional Chinese medicine). Processing A. kusnezoffii with a decoction of Terminalia chebula Retz. represents a characteristic and widely applied detoxification method in Mongolian medicine practice. This processing technique can reduce the toxicity of A. kusnezoffii . However, the underlying mechanisms remain insufficiently understood. Methods: Normal Sprague–Dawley (SD) rats were randomly divided into the control (Ctrl), raw A. kusnezoffii (SCW), and Terminalia chebula decoction-processed A. kusnezoffii (HZZCW) groups. The metabolic profiles of rat feces and intestinal microbiota were detected. Hematoxylin and eosin (H&E) staining and enzyme-linked immunosorbent assay (ELISA) were performed to verify the toxicity of A. kusnezoffii . Western blot (WB) and RT-qPCR were used to assess the expression of the PI3K/AKT signaling pathway. In addition, a collagen-induced arthritis (CIA) model was established to evaluate the anti-inflammatory effect. Results: Metabolomic analysis revealed that processing A. kusnezoffii with Terminalia chebula decoction significantly altered metabolic pathways, particularly those related to ubiquinone biosynthesis, porphyrin and pyrimidine metabolism. The results of 16S rDNA sequencing showed that the processed product markedly changed the α-diversity of the gut microbiota and reduced the relative abundance of Xanthomonas and Corynebacterium. Pharmacodynamic evaluation indicated that HZZCW reduced the toxicity of raw A. kusnezoffii by regulating the levels of malondialdehyde (MDA), lactate dehydrogenase (LDH), and superoxide dismutase (SOD), while activating the PI3K/AKT pathway to maintain its therapeutic efficacy. Furthermore, the CIA model confirmed its significant anti-inflammatory activity. Conclusion: Terminalia chebula Decoction-processed A. kusnezoffii may exert its pharmacodynamic effects through multiple mechanisms, including modulation of ubiquinone biosynthesis, porphyrin metabolism, and pyrimidine metabolism, regulation of the intestinal microbiota, and activation of the PI3K/AKT signaling pathway.

Therapeutic targeting of the CD47-SIRPα axis has emerged as a pivotal strategy for enhancing phagocytic clearance of tumor cells and overcoming immune evasion in solid malignancies, yet clinical outcomes with single-agent checkpoint blockade remain suboptimal due to limited activation of innate immunity and on-target toxicities. Administration of STING agonists is hampered by poor pharmacokinetics and nonspecific toxicity, highlighting the need for cell-selective delivery systems that can orchestrate immune activation and checkpoint inhibition. Herein, we conjugated cGAMP to an anti-SIRPα antibody, aiming to effectively reprogram the tumor immune microenvironment. Systemic administration of anti-SIRPα-cGAMP conjugate significantly inhibited tumor growth in a mouse model of colon adenocarcinoma and improved survival in vivo by simultaneously blocking the CD47-SIRPα "do not eat me" checkpoint and selectively delivering the STING agonist to myeloid cells. These findings suggest that targeted immunostimulatory ADCs may be an effective strategy for overcoming immune resistance in solid tumors.

Background: Brucella outer membrane proteins (Omps) are an important part of its cell wall and major virulence-related factors. Omp16 and Omp19 proteins are the advantageous antigens of Brucella and have been widely used in research on vaccines against brucellosis. As an emerging vaccine, the mRNA vaccine has unique advantages in the fight against intracellular parasitic bacteria. Methods: In this study, mRNA encoding the omp16 and omp19 genes of Brucella. melitensis (B. melitensis) was synthesized using in vitro transcription. The target mRNA was transfected into HEK 293T cells to evaluate protein expression levels and assess its immunogenicity. Finally, bioinformatic approaches were employed to analyze potential antigenic epitopes. Results: In this study, the successfully constructed recombinant plasmids pIVTRup-omp16 and pIVTRup-omp19 were utilized to synthesize omp16-mRNA and omp19-mRNA, each approximately 600 nt in length. Western blot analysis detected the expression of proteins with molecular weights of 16 kDa and 19 kDa in HEK 293T cells at 24 h post-transfection with mRNA. Purified rOmp16 and rOmp19 had good immunogenicity, which could specifically bind to serum antibodies of brucellosis patients. rOmp16 had stronger immunogenicity than rOmp19. Epitope prediction showed that Omp16 contained seven epitopes and Omp19 contained six epitopes. In addition, Omp16 and Omp19 could form stable complexes with target receptors. Simulated immunization with Omp16 and Omp19 proteins significantly activated both CD4⁺ and CD8⁺ T cells. Conclusions: The immunogenic proteins were successfully expressed in cells based on the mRNA fragments synthesized from omp16 and omp19 genes of B. melitensis, which was a preliminary exploration for the preparation of B. melitensis mRNA vaccine.

CpG oligodeoxynucleotides (CpG ODNs) are short, synthetically produced DNA molecules that serve as classic agonists of Toll-like receptor 9 (TLR9). They are known for their ability to combat infection, alleviate allergic responses, and inhibit tumor growth. However, free CpG is rapidly degraded by nucleases, distributes poorly, and accumulates minimally in tumors. To overcome these limitations, CpG was tethered to tumor-associated antigens or antibodies that engage Fcγ receptors on immune cells in previous studies. Here we redirected CpG toward myeloid cells via SIRPα, an innate immune checkpoint expressed in dendritic cells and macrophages that transmits "don't-eat-me" signals upon binding tumor-expressed CD47. The conjugate potently matured bone marrow-derived dendritic cells in vitro and, after simple intraperitoneal delivery, safely suppressed tumor growth in vivo. This myeloid-targeted CpG platform broadens the design space for TLR9 agonists and offers an off-the-shelf strategy for integrating innate and adaptive immunity against cancer.

Brain-derived neurotrophic factor (BDNF) plays an important role in synaptic development and plasticity. It is a promising therapeutic target for improving neurofunctional outcomes after traumatic brain injury (TBI). However, the delivery of BDNF faces several significant challenges including limited entry into the CNS due to blood-brain barrier (BBB), short half-life, and potential side effects. The use of viral vectors like AAV to deliver the BDNF gene directly to the brain has shown promise in animal models. However, issues with host immunogenicity and limited biodistribution remain. Herein, we report a successful restoration of cognitive function of a TBI mouse model by efficient delivery of BDNF mRNA loaded to a novel lipid nanoparticle (DA6 LNP). DA6 LNPs loaded with either luciferase mRNA or GFP mRNA were internalized by astrocytes and dose dependently expressed the corresponding protein. Two consecutive intravenous injections of DA6 LNPs loaded with BDNF mRNA to a TBI mouse model resulted in overexpression of BDNF in the brain and ameliorated cognitive impairment. Collectively, our data suggested that DA6 LNP is a promising carrier for CNS targeted delivery of therapeutic RNAs.

The twist operation of the π-skeleton would induce left or right handedness. Herein, we report two chirally twisted C₆₄ nanographenes designed by formally erasing two fused benzenes from a parent C₇₆ nanographene. These C₆₄ nanographenes possess two [6]helical units implemented around a central pyrene core in a different arrangement. The π-truncation pulls the emission bands hypsochromically from the red (C₇₆, λ_max = 612 nm) to green (C₆₄, λ_max = 505 and 506 nm) region. Curiously, the +/- sign of circularly polarized luminescence at the S₁ → S₀ transition was reversed between the two C₆₄ nanographenes despite them possessing the same handedness. This is attributed to the slight difference in the arrangement of the two transition dipole moments fluctuating at around 90°, as suggested computationally.

Purpose: Traditional Mongolian Medicine Batri-7 (BT-7) is the key Mongolian Medicine (MM) for bacterial enteritis. BT-7 is a well-known clinical MM due to its antibacterial properties. BT-7 contains plant-derived bioactive compounds, but its molecular mechanism of action remains unclear. This study explores BT-7’s antibacterial compounds and therapeutic mechanism in a Salmonella enteritis mouse model. Methods: The active components of BT-7 were detected by liquid chromatography-tandem mass spectrometry assay and identified by UPLC/Q-TOF-MS. An enteritis mouse model induced by Salmonella typhimurium was used in this study. Pathological analysis of small intestine was conducted with hematoxylin and eosin staining. The macrophage recruitment in model mice’s intestines was detected by flow cytometry. Simultaneously, the Minimum Inhibitory Concentration of BT-7 was evaluated against bacterial by microbroth dilution method, BT-7 regulation of Salmonella typhimurium gene was performed by RNA-Seq methods and verified by qRT-PCR. Results: In the LC-MS/MS assay, negative and positive-ion modes are identified for 511 and 699 compounds from BT-7, respectively. Of them, we found multiple antibacterial and anti inflammation compounds including chrysin, oroxylin A and luteolin. In vivo, we observed that treatment of mouse Salmonella enteritis with BT-7 decreases inflammation score and macrophages on intestinal tenue. In vitro, BT-7 presented the highest antibacterial activities against tested strains with MIC was 2– 4 mg/mL. Meanwhile, BT-7 significantly down regulated Salmonella infection genes. Conclusion: Twenty key anti-bacterial components were identified in the BT-7. In vivo experiment shows that orally administered BT-7 effectively reduce the inflammation of intestine in model of Salmonella-induced mouse enteritis by down regulating the infection-related virulence genes of Salmonella. Through this study, we discovered the mechanism of BT-7’s dual action on the host and pathogenic bacteria. This gives inspiration for anti-infective disease research in traditional medicine and also proves that traditional medicines still have good prospects for treating infectious diseases.