The aim of this study is to evaluate the clinical efficacy of Fufang Zaofan Pills in individuals diagnosed with lower-risk myelodysplastic syndrome (MDS) and to assess associated changes in peripheral blood dendritic cell (DC) levels. A cohort of 60 individuals with newly diagnosed lower-risk MDS were enrolled at the Department of Hematology, Jiading Central Hospital, affiliated with Shanghai University of Medicine & Health Sciences between February 2022 and July 2023. Participants were randomly allocated into two groups: an integrated Chinese-Western medicine group (n = 40) and a Western medicine group (n = 20). Both groups received standard Western medical therapy as per clinical guidelines, while the integrated group received additional oral administration of Fufang Zaofan Pills. Following the treatment course, clinical outcomes were compared between groups, including Western medicine efficacy, changes in traditional Chinese medicine (TCM) syndrome scores, and peripheral blood DC levels before and after treatment. At baseline, lower-risk MDS patients exhibited significantly elevated levels of DC and mDC compared to healthy controls (p < 0.05). The objective response rate (ORR) in the integrated Chinese-Western medicine group was 47.5%, which was significantly higher than the 20% observed in the Western medicine group (p < 0.05). The total effective rate for TCM syndrome scores was also significantly greater in the integrated group (80% vs. 60%, p < 0.05). Post-treatment analysis indicated significant changes in DC percentage, absolute count, and mDC-related indices in the integrated group compared to baseline values (p < 0.05), with intergroup comparisons demonstrating statistically significant differences in favor of the integrated therapy (p < 0.05). The addition of Fufang Zaofan Pills to standard treatment appears to enhance clinical efficacy in lower-risk MDS and is associated with modulation of peripheral blood DC levels. These preliminary findings contribute evidence supporting further investigation into the integration of TCM and Western medical approaches in the management of lower-risk MDS, particularly regarding their immunomodulatory potential.

This study aimed to investigate the therapeutic potential of Tanshinone IIA (Tan-IIA) in alleviating chronic endometritis (CE), focusing on its role in mitigating mitochondrial dysfunction and suppressing neutrophil extracellular traps (NETs) formation. HEnEpCs were stimulated with lipopolysaccharide (LPS) to establish an inflammatory model. Application of Tan-IIA as a pretreatment was followed by a comprehensive assessment of mitochondrial function, including JC-1 staining, adenosine triphosphate (ATP) content, reactive oxygen species (ROS) levels, and transmission electron microscopy. A co-culture system with human neutrophils was used to evaluate NETs formation. Molecular mechanisms were probed using real-time quantitative PCR (RT-qPCR), Western blot, chromatin immunoprecipitation (ChIP), and immunofluorescence. A rat model of LPS-induced endometritis was used to validate the effects of Tan-IIA on uterine histopathology, NETs formation, and inflammatory cytokine levels. Tan-IIA significantly ameliorated LPS-induced mitochondrial damage, restored membrane potential, reduced ROS production, and increased ATP levels in HEnEpCs. DRAK2 was elevated in CE patient plasma and LPS-stimulated cells, of which overexpression antagonized protective effects mediated by Tan-IIA. Furthermore, Tan-IIA inhibited NETs formation in co-culture systems, an effect mediated through suppression of the lysine demethylase 3A (KDM3A)-histone H3 lysine 9 dimethylation (H3K9me2)-hypoxia-inducible factor 1-alpha (HIF-1A)-solute carrier family 2 member 3 (SLC2A3) epigenetic-metabolic axis. Tan-IIA reduced neutrophil infiltration, NETs formation, and pro-inflammatory cytokine levels to improve endometrial architecture in LPS-induced rats. Tan-IIA attenuates CE progression by targeting DRAK2 to restore mitochondrial function and inhibiting NETs formation via the KDM3A/SLC2A3 pathway, implying Tan-IIA as a promising multi-target agent for CE therapy.

Human umbilical cord mesenchymal stem cells-derived exosomes (UC-MSCs-exos) have demonstrated considerable potential in osteoporosis treatment. This study explored the underlying mechanisms of UC-MSCs-exos in this context. Osteoporosis cell models were established by treating RAW264.7 cells with RANKL, followed by incubation with UC-MSCs-exos. The effects on osteoclast differentiation were assessed via Western blot and TRAP staining. Transfection of HECTD4, METTL3, and RANK was performed to investigate the regulatory mechanism of UC-MSCs-exos on osteoclast differentiation. MeRIP-qPCR analyzed the impact of METTL3 on RANK m6A modification, and ubiquitination assays examined the influence of HECTD4 on METTL3 ubiquitination. Treatment with UC-MSCs-exos inhibited osteoclast differentiation in the osteoporosis models by decreasing the number of mature osteoclasts and markers such as RANK, ACP5, and CTSK. UC-MSCs-exos also reduced METTL3 levels in these models. When METTL3 was overexpressed in the presence of UC-MSCs-exos, osteoclast differentiation was enhanced. METTL3 promoted RANK protein stability by facilitating its m6A modification. In UC-MSCs-exos-treated models, RANK silencing reversed the effect of METTL3 overexpression on osteoclast differentiation. UC-MSCs-exos were found to contain abundant HECTD4, which promoted METTL3 degradation via ubiquitination. Silencing of HECTD4 promoted osteoclast differentiation in the presence of UC-MSCs-exos, but this was reversed by METTL3 silencing. HECTD4 from UC-MSCs-exos attenuated osteoclast differentiation in osteoporosis by inhibiting RANK m6A modification through METTL3 ubiquitination. These findings suggest that UC-MSCs-exos offer a promising strategy for osteoporosis treatment.

Reduced response to targeted therapy in human epidermal growth factor receptor 2 (HER2)-positive breast cancer underscores the need for novel therapeutic strategies and molecular targets. Cluster of differentiation 24 (CD24), an oncogenic molecule closely associated with aggressive tumor phenotypes, was investigated for its expression in HER2-positive breast cancer and its potential as a candidate target for combination therapy. Herein, we explored the expression of CD24 in breast cancer (BC) and its prognostic significance. In HER2-positive BC cells, CD24 was knocked down to assess its impacts on tumor biological behaviors and synergistic effects with trastuzumab. Furthermore, the underlying mechanism of CD24 was elucidated using a series of experimental assays, including Western blotting (WB), immunohistochemistry (IHC), terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay, and mRFP-GFP-LC3 dual-fluorescence autophagy assay. In vivo experiments were conducted to evaluate the therapeutic efficacy of CD24 inhibition combined with trastuzumab. CD24 was highly expressed in HER2 + breast cancer and associated with poor prognosis. CD24 knockdown significantly suppressed the proliferation, migration, and invasion of HER2-positive BC cells. Moreover, CD24 depletion promoted apoptosis and autophagy in HER2-positive breast cancer, accompanied by the inactivation of the Akt/mammalian target of rapamycin (mTOR) signaling pathway. In vivo data demonstrated that the combination regimen of CD24 knockdown and trastuzumab markedly reduced tumor burden. CD24 silencing modulates autophagy and induces apoptosis via Akt/mTOR inhibition, and its combination with trastuzumab exerts potent antitumor effects, offering a promising strategy for HER2 + breast cancer.

Granulomatous lobular mastitis (GLM) is a chronic, refractory inflammatory condition of the breast with an unclear etiology. Emerging evidence suggests a potential role of pyroptosis in the pathogenesis of GLM. This study investigated the therapeutic effects of Tuolitounong Decoction (TLTND) on pyroptosis in both in vitro and in vivo models of GLM. An in vitro GLM model was established using MCF10A mammary epithelial cells exposed to tissue homogenates derived from GLM-affected human breast tissue. Cells were treated with TLTND at varying concentrations, with or without the pyroptosis inhibitor necrosulfonamide. Pyroptosis-associated proteins, including caspase-1, gasdermin D (GSDMD), lipopolysaccharide-binding protein (LBP), and NLR family pyrin domain containing 3 (NLRP3), were assessed using immunofluorescence and western blotting. Cell viability was evaluated using the Cell Counting Kit-8 (CCK-8) assay. For in vivo analysis, a rat GLM model was induced by injecting a combination of human GLM tissue homogenate supernatant and Freund's complete adjuvant into the third and fourth pairs of mammary glands in female Sprague Dawley rats. TLTND was administered via daily oral gavage for 21 days. Post-treatment evaluations included histopathological assessment, expression of estrogen receptor (ER) and progesterone receptor (PR), and pyroptosis-related biomarkers in mammary gland tissue. Exposure to GLM homogenate successfully induced pyroptosis-related pathological features in MCF10A cells, whereas serotonin (5-HT) inhibition showed no significant effect. TLTND treatment demonstrated dose- and time-dependent effects, with enhanced therapeutic efficacy at 10 mg/mL after 48 h (P = 0.009). Of note, TLTND significantly reduced the protein expression levels of key pyroptosis markers (cleaved caspase-1, n-GSDMD, cleaved IL-18 and cleaved IL-1β) by 52.6%, 91.0%, 78.1% and 87.1%, respectively, compared to the model group in rat mammary tissue. Co-treatment with TLTND and necrosulfonamide significantly downregulated the expression of caspase-1, GSDMD, LBP, and NLRP3, while increasing cell viability. In vivo, both prednisone acetate and TLTND ameliorated histopathological features of GLM. Notably, TLTND promoted localized abscess maturation and increased ER and PR expression, while reducing levels of interleukin-1β and GSDMD-N in mammary tissue. TLTND demonstrated therapeutic potential in reducing pyroptosis in mammary epithelial cells and modulating inflammation in granulomatous lobular mastitis, partly by downregulating the caspase-1/GSDMD-mediated pyroptosis signaling pathway. These findings support further investigation of TLTND as a complementary treatment approach for GLM and provide insight into its underlying mechanisms of action.

Modified citrus pectin (MCP), a polysaccharide from citrus fruits, modulates galectin-3 (Gal-3) and immune responses. Although MCP exhibits notable immunomodulatory effects, its role in drug-induced splenic and systemic toxicity remains unexplored. This study investigated the effects of MCP modulation on splenic immune remodeling and galectin expression in a rat model of cisplatin-induced toxicity. Wistar rats were divided into four groups (n = 5 animals/group): control (SHAM), MCP (100mg/kg/day for 7 days), cisplatin (CIS, 10mg/kg/day for 3 days), and MCP + CIS. Spleens were collected 6h after the final cisplatin dose. Cisplatin induced splenic structural disorganization and selectively reduced nitric oxide levels without broadly affecting antioxidant enzymes. Cisplatin treatment was associated with increased CD3⁺ T cell labeling and enhanced tissue expression of Gal-1, -3, and - 9. MCP administration did not restore splenic architecture but promoted increased hemosiderin deposition in the red pulp, suggestive of enhanced erythrophagocytosis and altered iron handling, and markedly reduced CD3⁺ T cell immunoreactivity. MCP associated with cisplatin also reduced Gal-1 and Gal-3 levels and altered the relationship between galectins and splenic immune cell populations. Correlation analyses revealed positive associations between Gal-1, -3, and - 9 and CD68⁺ macrophages, as well as a selective association between Gal-3 and CD3⁺ T cells, exclusively in MCP + CIS animals. MCP does not mitigate cisplatin-induced splenic damage but alters immune cell distribution and galectin-related responses during cisplatin exposure.

Postmenopausal osteoporosis (PMOP) is a skeletal disorder marked by progressive bone mineral density decline and increased susceptibility to fractures. Accumulating evidence indicates that coupled osteogenesis and angiogenesis are indispensable for bone remodeling and repair. This study investigated the therapeutic potential of dendrobine in concurrently modulating osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) and angiogenic activity in the context of PMOP. BMSCs were isolated from Sprague-Dawley rats and rigorously characterized prior to experimentation. Cell viability was quantified using CCK-8 assays, and apoptosis was assessed via flow cytometry. To evaluate angiogenesis, tube formation capacity was measured following coculture of BMSCs and human umbilical vein endothelial cells (HUVECs), and protein levels of angiogenic factors (VEGF, MMP2, and MMP9) were quantified by western blotting. Osteogenic differentiation was assessed by Alizarin red staining for mineralized nodule formation, ALP staining, and western blotting of osteogenic markers (Runx2, OCN, and OPN). Following PMOP modeling via bilateral ovariectomy (OVX), H&E staining and micro-computed tomography were conducted. Western blotting was performed to measure protein levels of core Hippo pathway components (Mst1, Lats1, Yap1, and Taz) in femoral bone tissue. Results showed that dendrobine significantly promoted BMSC viability and suppressed apoptosis in a dose-dependent manner. In BMSC-HUVEC cocultures, dendrobine markedly augmented tube formation and upregulated VEGF, MMP2, and MMP9. Concurrently, dendrobine enhanced osteogenic differentiation, as evidenced by increased ALP activity, enhanced mineralization, and elevated protein levels of Runx2, OCN, and OPN. In OVX rats, dendrobine treatment ameliorated trabecular bone loss and restored bone parameters toward sham levels. Consistently, dendrobine elevated the expression of osteogenic and angiogenic proteins in vivo. Moreover, dendrobine suppressed Mst1 and Lats1 expression while promoting Yap1 and Taz expression in OVX rats. In conclusion, dendrobine alleviates PMOP by coordinately enhancing osteogenesis and angiogenesis through inhibition of the Hippo pathway and subsequent activation of the Yap1/Taz signaling.

Unexplained miscarriage (UM) is a distressing pregnancy disorder often associated with impaired trophoblast function. This study aimed to investigate the role of C-type lectin domain family 14 member A (CLEC14A) in the pathogenesis of UM and to elucidate the underlying molecular mechanisms. Transcriptomic data from UM and healthy placental tissues (GSE123719) were analyzed. CLEC14A expression was validated in clinical chorionic villi samples (n = 10 per group) by immunohistochemistry, RT-qPCR, and Western blot. Gain-of-function studies were performed in HTR-8/SVneo trophoblast cells using lentiviral-mediated CLEC14A overexpression. Functional impacts on migration, invasion, and tube formation were assessed. RNA sequencing was used to identify dysregulated pathways, and key findings were confirmed by ELISA and Western blot. Rescue experiments were conducted using the adenylate cyclase inhibitor SQ22536. CLEC14A was significantly upregulated at both the mRNA and protein levels in villous tissues from UM patients compared to controls. In vitro, CLEC14A overexpression markedly suppressed trophoblast migration, invasion, and endothelial-like tube formation. Transcriptomic and mechanistic analyses revealed that CLEC14A overexpression activated the cAMP signaling pathway, leading to increased intracellular cAMP levels and Protein Kinase A (PKA) phosphorylation, while concurrently attenuating extracellular signal-regulated kinase (ERK) phosphorylation. The functional deficits and signaling perturbations induced by CLEC14A were effectively reversed by inhibition of cAMP synthesis. Our findings demonstrate that aberrant upregulation of CLEC14A impairs trophoblast function by activating the cAMP-PKA pathway and suppressing ERK signaling, thereby contributing to the pathogenesis of unexplained miscarriage. CLEC14A and its downstream signaling axis represent potential diagnostic and therapeutic targets for this condition.

Sleep amount and quality are important to maintain health, light exposure transported to the pineal gland via suprachiasmatic nucleus pineal pathway, and this affect melatonin synthesis. Melatonin exerts its action by receptors called MT1, MT2 that found in peripheral tissues including pancreas, which affect its endocrine function. In this study we investigate the effect of type of sleep disturbance on the histological structure and the expression of melatonin receptor (MTNR1B) in the pancreas. A sample of 45 adult healthy male rats had free access to water and feeding, divided into 3 groups (15 in each group): the control group had normal 24 h diurnal variation, group A subjected to interruption of sleep by light exposure for 2 h at three intervals, and group B subjected to a reduction in sleep duration by 7 h. This experiment continued for 14 days, animals were scarified by euthanasia by cervical dislocation, pancreatic tissue prepared for paraffin blocks and stained by H&E, and IHC for MTNR1B. Histomorhometric analysis done by image J (V 1.54), immunohistochemical assessment done by Aprioscope image analysis software 12.4.6.5003) mean total positivity of expression was selected, statistical analysis done by SSPS (25 V) and Tukeyꞌs test. Histological evaluation revealed changes in sleep disturbance groups including fat deposition, vascular dilatation, and apoptotic changes for group of cells within islets of Langerhans which showed significant reduction in their count in sleep disturbance groups compared to control, and non-significant change in their area across the groups, but significant reduction in area in sleep deprived group compared to sleep interruption. Immunohistochemical expression of MTNR1B showed it only expressed within the area of islets of Langerhans, with non-significant increase in expression in sleep disturbed groups, being more in sleep deprived group. This study showed that the pancreas is affected by sleep disturbance patterns, mainly its endocrine part (islets of Langerhans), with study expression of melatonin receptor within them. This may suggest a role of melatonin in maintaining pancreatic ꞵ cells.

This study aimed to investigate the analgesic effects of lidocaine (Lido) on diabetic neuropathic pain (DNP), a common complication of diabetes, and to elucidate the underlying mechanism. A DNP mouse model was established by a single streptozotocin injection at 200 mg/kg. Two weeks later, mice received intraperitoneal infusion with 2% Lido for 7 consecutive days. The analgesic effect of Lido was assessed by pain behavioral tests. Astrocyte activation, pro-inflammatory cytokine levels, and CXCL13 and CXCR5 expression were examined in mechanistic studies. Lido significantly improved mechanical allodynia and thermal hyperalgesia in DNP mice. Lido reduced spinal astrocyte activation and pro-inflammatory cytokine levels in DNP mice. It also inhibited CXCL13 and CXCR5 expression, with their upregulation diminishing Lido's analgesic effects. Lido alleviates DNP by inhibiting spinal astrocyte activation via modulation of the CXCL13/CXCR5 axis.