To explore the similarities and differences in the biological phenotypes and cytotoxicity of resident and expanded natural killer cells derived from mononuclear cells of human bone marrow between healthy donors (HD) and patients with B-cell acute lymphoblastic leukemia (B-ALL) .

Bone marrow blood of HD and B-ALL was collected, and mononuclear cells were enriched by density gradient centrifugation. Flow cytometry was used to analyze the phenotypes of resident NK cells (HD-NK and BALL-NK) in the two types of blood. Then, with the aid of our well-established 14-day "3ILs"-based (short for IL-2, IL-15, IL-18) culture system for in vitro high-efficient activation and expansion of NK cells, we conducted cell counting, flow cytometry analysis and in vitro co-culture killing tests to compare the phenotypes and cytotoxicity of expanded HD-NK cells (eHD-NK) and expanded BALL-NK cells (eBALL-NK) in bone marrow blood. Comparisons between two groups were performed using independent samples t-test. Comparisons among the four groups (HD-NK, eHD-NK, BALL-NK, and eBALL-NK) were conducted using two-way repeated measures ANOVA, followed by Bonferroni's post hoc test for pairwise comparisons.

Compared with the healthy donor (HD) group, the proportion of total CD3^-CD56⁺ NK cells in the non-activated, expanded resident NK cell population from patients with B-cell acute lymphoblastic leukemia (B-ALL) was significantly lower [ (4.21 ± 0.92) % vs (14.25 ± 1.15) %]. In addition, the frequencies of activated NK cell subsets were reduced, including CD16⁺ NK cells [ (46.82 ± 2.96) % vs (67.87 ± 2.12) %], NKG2D⁺ NK cells [ (22.45 ± 2.12) %vs (50.82 ± 5.65) %], and NKp46⁺ NK cells [ (7.66 ± 1.73) % vs (17.27 ± 1.75) %]. Following in vitro activation and expansion, the proportion of total NK cells was significantly increased in both the expanded HD-NK (eHD-NK) group and the expanded B-ALL-derived NK (eBALL-NK) group [ (61.52 ± 3.18) % vs (14.25 ± 1.15) %]; (24.63 ± 2.07) % vs (4.21 ± 0.92) %]. Moreover, the eHD-NK group exhibited higher proportions of total NK cells [ (61.52 ± 3.18) % vs (24.63 ± 2.07) %] and multiple activated NK cell subsets than the eBALL-NK group, including NKG2D⁺ NK cells [ (80.63 ± 2.03) % vs (57.83 ± 8.55) %], CD25⁺ NK cells [ (37.45 ± 3.21) %vs (20.90 ± 5.15) %], and NKp46⁺ NK cells [ (27.23 ± 2.30) % vs (9.51 ± 0.98) %]. Cell viability analysis demonstrated that the efficiency of in vitro activation and expansion of total NK cells was higher in the HD group than in the B-ALL group. Compared with the eHD-NK group, the eBALL-NK group showed significantly higher proportions of apoptotic cells, including Annexin V⁺7-AAD- cells [ (4.50 ± 0.35) % vs (2.72 ± 0.43) %] and Annexin V⁺ cells [ (5.13 ± 0.62) % vs (3.29 ± 0.58) %], while no significant differences in cell cycle distribution were observed between the two groups. In vitro cytotoxicity co-culture assays revealed that eHD-NK cells exhibited stronger cytotoxic activity against the tumor cell lines Nalm6 and U937 compared with eBALL-NK cells.

Compared with HDs, bone marrow–derived resident and activated NK cells from B-ALL patients exhibited reduced proportions, diminished activated subpopulation content, impaired in vitro activation capacity, decreased cellular viability, and attenuated cytotoxicity. This study provides valuable reference for subsequent investigations into the underlying pathogenic mechanisms and the development of clinical therapeutic strategies.

Exploring the ameliorative effects of blueberry extract on atopic dermatitis (AD) and elucidating its mechanisms of action through changes in skin microbiota, restoration of barrier function, and improvement of Th2-type inflammation.

The AD mouse model was induced by topical application of the vitamin D3 analog calcipotriol (MC903). Ear thickness was measured in a double-blind manner on days 1, 3, 5, 7, 10, 12, and 14, and scratching episodes within 30 minutes were recorded on day 14. Ear tissue samples were collected for hematoxylin-eosin (HE) staining and epidermal thickness measurement. The mRNA expression of inflammatory cytokines (IL-33, IL-22, TSLP, TNF-α, IL-6, and IL-1β) was detected by RT-qPCR. 16S rRNA sequencing was used to analyze the effect of blueberry extract on the microbial community in AD mice. A cellular model was established by stimulating human keratinocytes (HaCaT) with lipopolysaccharide (LPS, 20 μg/mL). The effect of different concentrations of blueberry extract on cell viability was assessed using the CCK-8 assay. Expression of IL-33, TSLP, and IL-4Rα proteins were detected by Western blot. The composition of blueberry extract was identified by ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). Comparisons between two groups were performed using the independent samples t-test. Comparisons among multiple groups were conducted using either one-way ANOVA or repeated-measures ANOVA, as appropriate. Post hoc pairwise comparisons were carried out with Tukey's test.

Compared with the model group, blueberry extract significantly alleviated ear swelling in AD mice, reduced scratching frequency [ (9.00 ± 0.89) vs (14.67 ± 2.06) times] and epidermal thickness [ (37.78 ± 2.68) vs (54.90 ± 10.09) μm] (all P < 0.05). Blueberry extract downregulated the expression of Th2 inflammatory cytokines and pro-inflammatory factors mRNA. Compared with the model group, the expression levels of IL-33 (0.95 ± 0.18 vs 1.36 ± 0.23), TSLP (27.12 ± 4.73 vs 39.09 ± 4.61), IL-22 (1.98 ± 0.45 vs 3.43 ± 0.36), and IL-1β (4.47 ± 1.44 vs 20.26 ± 3.30) mRNA were decreased in treatment group (P < 0.05). 16S rRNA analysis revealed that blueberry extract mitigated microbial community dysbiosis and loss of diversity, inhibiting the proliferation of pathogenic bacteria such as Streptococcus, Staphylococcus, and Enterobacter. In the cellular experiments, compared with the LPS-induced model group, treatment with 80 μmol/L blueberry extract significantly reduced the protein expression of IL-4Rα (1.64 ± 0.31 vs 5.90 ± 0.24), IL-33 (0.90 ± 0.15 vs 4.27 ± 0.41), and TSLP (0.15 ± 0.07 vs 5.61 ± 0.40), as well as the mRNA expression of IL-4Rα (2.57 ± 0.54 vs 13.20 ± 3.43), IL-33 (2.00 ± 0.44 vs 5.40 ± 1.24), and TSLP (1.47 ± 0.06 vs 4.04 ± 0.21) (all P < 0.05). Further mass spectrometry identification indicated that delphinidin might be the core active component in blueberry extract.

Blueberry extract significantly alleviated symptoms in the AD mouse model by modulating the skin microbiota, restoring barrier function, suppressing Th2-type inflammatory responses, and reducing inflammatory cytokine expression in HaCaT cells, with delphinidin identified as its potential core active constituent.

The neddylation process covalently conjugates neural precursor cell expressed developmentally downregulated 8 (NEDD8) to substrate proteins through a continuous enzymatic cascade reaction, which affects the stability, conformation, and function of substrate proteins, thereby regulating numerous downstream biological processes. The neddylation process is associated with the progression and prognosis of various types of tumors. As the only currently known activating enzyme in the neddylation process, NEDD8-activating enzyme (NAE) is an important target for the development of anti-cancer drugs. MLN4924, a selective NAE inhibitor, has good therapeutic effects on various types of solid tumors and hematological malignancies. Currently, 41 clinical trials have entered phases Ⅱ/Ⅲ, indicating promising application prospects. The drug resistance of tumor cells is an important cause of treatment failure in cancer patients. Once tumor cells acquire drug resistance, it will significantly affect the prognosis of patients. This article reviews the current research progress on the resistance mechanism of MLN4924 and the measures to address the problem of resistance to it.

Renal fibrosis is a multifactorial-driven pathological process involving intrarenal inflammation, cellular proliferation, and excessive deposition of collagen and fibronectin. Without intervention, this process can progress to chronic kidney disease (CKD) and may ultimately lead to end-stage renal disease. Stem cell-derived exosomes represent an emerging therapeutic strategy for renal fibrosis. Functioning as nanoscale extracellular vesicles (EVs), exosomes carry bioactive molecules that can be taken up by local or distal cells. They serve as mediators of intercellular communication while also delivering therapeutic payloads (e.g., microRNAs, proteins). In animal models of kidney disease, stem cell-derived exosomes have been demonstrated to effectively attenuate inflammation, suppress fibroblast activation, and reduce collagen production in the renal interstitium, thereby delaying fibrosis progression. This review summarizes the mechanisms underlying the antifibrotic effects of exosomes derived from various stem cell sources used as therapeutic agents for renal fibrosis. It also outlines future translational research directions and strategies aimed at enhancing the therapeutic efficacy of stem cell-derived exosomes.

Poor graft function (PGF) following allogeneic hematopoietic stem cell transplantation (allo-HSCT) is a severe post-transplant complication affecting patient prognosis, which often leads to infections and an elevated risk of hemorrhagic events. The pathogenesis of PGF is multifactorial, involving mechanisms such as functional impairment of donor-derived stem cells, injury to the bone marrow microenvironment, and dysregulation of immune homeostasis. Current therapeutic strategies for PGF include mesenchymal stem cell infusions, CD34⁺ cell reinfusion, secondary transplantation as well as drug intervention such as statins and hematopoietic growth factors, repairing the microenvironment and promoting hematopoietic reconstitution. This article reviews the recent advances in understanding the pathophysiological mechanisms and therapeutic management of PGF.

Diabetic nephropathy (DN) is the most common microvascular complications of diabetes and serves as a leading cause of end-stage renal disease, which is caused by complex mechanisms, including cellular damage induced by hyperglycemia, dysregulated inflammatory and immune responses, renal fibrosis, and vascular injury. Although conventional treatments can delay the progress of the disease, they often fail to effectively halt the irreversible deterioration of renal function. Consequently, exploring practical and effective therapeutic strategies is a key issue that needs to be urgently addressed. In recent years, mesenchymal stem cells (MSCs) and their exosomes have garnered significant research interest due to their capacities for self-renewal and multi-lineage differentiation. Preclinical studies indicate that MSCs exhibit potential in restoring renal structure and function through various mechanisms, such as cytoprotection, modulation of inflammation and immune responses, inhibition of fibrosis, epigenetic regulation, and regulation of the gut–kidney axis. Multiple administration routes, including intravenous, renal arterial, and local injection, have demonstrated considerable therapeutic efficacy. Nonetheless, clinical research remains in the nascent stages, with only small-scale trials and case reports indicating that the MSCs have certain potential for renal protection. Their long-term safety, standardized treatment regimens, and the efficacy differences among MSCs from different sources still require further research and verification.

Mesenchymal stem cells (MSCs) have shown broad application prospects in the field of regenerative medicine due to their strong self-renewal ability, low immunogenicity and multi-directional differentiation potential. However, during the in vitro expansion, due to the differences between the culture environment and the physiological state in vivo, MSCs are prone to replicative senescence, which is manifested as slowed proliferation ability, reduced differentiation capacity and weakened function, thereby limiting their clinical transformation and application. Recently, significant efforts have been made to develop effective strategies to delay MSC aging and have made significant progress in gene modification, optimization of culture systems, and application of biomaterials. This review highlights recent advances in approaches to delay senescence in MSCs, aiming to provide a theoretical basis for improving the quality of in vitro culture of MSCs and maintaining their functional characteristics and broaden the scope of MSC applications in tissue engineering and cell-based therapies.