ObjectiveTo investigate whether Huanglian Jiedutang can inhibit neutrophil infiltration in the brains of middle cerebral artery occlusion （MCAO） mice by regulating the expression of neutrophil-related chemokines in exosomes， thereby achieving therapeutic effects. MethodsA total of 130 male specific pathogen-free （SPF） C57BL/6J mice were randomly divided into four groups： Sham-operated group， MCAO model group， Huanglian Jiedutang group （6 g·kg^-1）， and Ginaton group （21.6 mg·kg^-1）， with 10 mice in the Ginaton group and 40 mice in each of the remaining three groups. Mice in the Huanglian Jiedutang group and the Ginaton group were administered the corresponding drugs by oral gavage once daily at a volume of 0.15 mL·（10 g）^-1 for 7 consecutive days， while the sham-operated and model groups received an equal volume of saline via the same route. After 7 days， MCAO surgery was performed. The distal and proximal ends of the right common carotid artery （CCA） were ligated， a small incision was made between the two ligatures， and a silicone rubber-coated monofilament with a rounded tip was inserted into the lumen to occlude the CCA. The filament was left in place for 1 h to establish a focal cerebral ischemia model. At 24 h after modeling， mice were evaluated. Neurological function was assessed using the Longa score. Cerebral infarct volume was measured by 2，3，5-triphenyltetrazolium chloride （TTC） staining. Cerebral blood flow was observed by laser speckle imaging. Hematoxylin and eosin （HE） staining and Nissl staining were used to observe pathological changes in brain tissues. Exosomes were isolated from mouse plasma and brain tissues by ultracentrifugation and molecular size exclusion and identified by electron microscopy， particle size analysis， and protein blotting. Long-chain RNA libraries of exosomes were constructed and sequenced. Real-time quantitative reverse transcription polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of inflammatory factors and neutrophil-related chemokines in exosomes from plasma and brain tissues of each group. Enzyme-linked immunosorbent assay （ELISA） was used to detect the protein expression of inflammatory factors and neutrophil-related chemokines in exosomes from brain tissues of each group. Immunohistochemistry was used to detect the expression of the neutrophil-specific protein myeloperoxidase （MPO） in the brains of mice in each group. ResultsCompared with the sham-operated group， the model group showed decreased neurological function scores （P<0.01）， obvious cerebral infarction （P<0.01）， reduced cerebral blood flow （P<0.01）， neuronal necrosis in the brain， and decreased numbers of Nissl bodies （P<0.01）. The mRNA expression levels of IL-1β， MPO， CXCL1， CXCL2， CXCL3， CXCL10， CCL2， and CCL3 in exosomes from plasma and brain tissues were significantly increased （P<0.05， P<0.01）. The protein expression levels of IL-1β， MPO， CXCL2， and CXCL10 in exosomes from brain tissues were increased （P<0.05， P<0.01）， and MPO-positive rates and mean optical density values in brain tissues were elevated （P<0.01）. Compared with the model group， the Huanglian Jiedutang group and the Ginaton group showed increased neurological function scores （P<0.05）， reduced cerebral infarct volume （P<0.01）， restored cerebral blood flow （P<0.01）， reduced necrotic cells in the brain， and increased numbers of Nissl bodies （P<0.01）. In the Huanglian Jiedutang group， the mRNA expression levels of IL-1β， MPO， CXCL1， CXCL2， CXCL3， CXCL10， CCL2， and CCL3 in exosomes from plasma and brain tissues were decreased （P<0.05， P<0.01）. The protein expression levels of IL-1β， MPO， CXCL2， and CXCL10 in exosomes from brain tissues were reduced （P<0.05， P<0.01）， and MPO-positive rates and mean optical density values in brain tissues were decreased （P<0.01）. ConclusionHuanglian Jiedutang can effectively regulate the expression of neutrophil-related chemokines in exosomes from plasma and brain tissues of MCAO mice， thereby reducing neutrophil infiltration in the brain and achieving therapeutic effects.

Background 900 MHz radiofrequency radiation (RF) is a commonly used frequency in modern wireless communication devices, and its potential health effects have drawn much attention, especially its impact on bone metabolism, which has not been fully clarified. Objective To investigate the effects of 900 MHz RF on the bone tissue and osteoblast senescence of mice, as well as the dose-effect relationship. Methods In vivo, 3-month-old female C57BL/6 mice were divided into five groups (n=10): sham exposure, low-dose RF (50 μW·cm⁻²), medium-dose RF (150 μW·cm⁻²), high-dose RF (450 μW·cm⁻²), and D-galactose positive control (D-gal). Treatments were administered for 4 h per day for 28 d. Bone mineral density (BMD) and microstructure, including bone volume (BV), tissue volume (TV), bone volume fraction (BV/TV), trabecular number (Tb.N), trabecular separation (Tb.Sp), and trabecular thickness (Tb.Th), were assessed by Micro-CT; bone morphology was examined after hematoxylin and eosin (HE) staining; osteoprotegerin (OPG) and receptor activator of nuclear factor kappa-κΒ ligand (RANKL) expression was detected by immunohistochemistry; serum OPG, tartrate-resistant acid phosphatase 5b (TRACP-5b), plasminogen activator inhibitor-1 (PAI-1), interleukin-6 (IL-6), and C-X-C motif chemokine ligand 15 (CXCL15) levels were measured by enzyme-linked immunosorbent assay (ELISA); mRNA expression of Tp53, Cdkn1a, and Cdkn2a in bone tissue was analyzed by reverse transcription polymerase chain reaction (RT-PCR). In vitro, MC3T3-E1 pre-osteoblasts were grouped into sham, low-dose RF (50 μW·cm⁻²), medium-dose RF (150 μW·cm⁻²), high-dose RF (450 μW·cm⁻²), and H₂O₂ control, groups, and were exposed for 4 h per day for 5 d. Cell morphology was observed by microscopy; viability was tested by cell counting kit-8 (CCK-8); senescence was evaluated by senescence-associated β-galactosidase (SA-β-gal) staining; P53 and P21 protein expression was detected by Western blot; Tp53 and Cdkn1a mRNA levels were measured by RT-PCR. Results In vivo, RF at each dose significantly reduced the BMD of the mice's femurs and the bone microstructure parameters, such as BV/TV, Tb.N, and Tb.Th (P<0.05). Among them, Tb.Sp only increased in the 150 μW·cm⁻² RF group (P<0.05), with a looser bone network; fewer, sparser trabeculae and increased marrow fat were observed after HE staining; down-regulated OPG and up-regulated RANKL expression levels were observed by immunohistochemistry; the ELISA test revealed that the serum OPG levels in the 150 μW·cm⁻² RF group and the 450 μW·cm⁻² RF group of mice were significantly decreased (P<0.05), while the indicator in the 50 μW·cm⁻² RF group showed a decreasing trend but the difference was not statistically significant (P>0.05), TRACP-5b rose, and PAI-1, IL-6, and CXCL15 levels increased (P<0.05); the RT-PCR results showed thatTp53, Cdkn1a, and Cdkn2a mRNA expression was upregulated (P<0.05). In vitro, radiofrequency radiation induced cell flattening, reduced viability (P<0.05), increased SA-β-gal-positive cells (P<0.05), and upregulated P53, P21, Tp53, and Cdkn1a expression (P<0.05). Conclusion 900 MHz RF disrupts bone metabolism in mice by inhibiting bone formation, promoting resorption, and inducing osteoblast senescence, accelerating bone aging. The 150 μW·cm⁻² RF dose exhibits the most pronounced effect, reflecting a nonlinear “window effect,” highlighting potential health risks.

ObjectiveTo investigate whether Huanglian Jiedutang can inhibit neutrophil infiltration in the brains of middle cerebral artery occlusion （MCAO） mice by regulating the expression of neutrophil-related chemokines in exosomes， thereby achieving therapeutic effects. MethodsA total of 130 male specific pathogen-free （SPF） C57BL/6J mice were randomly divided into four groups： Sham-operated group， MCAO model group， Huanglian Jiedutang group （6 g·kg^-1）， and Ginaton group （21.6 mg·kg^-1）， with 10 mice in the Ginaton group and 40 mice in each of the remaining three groups. Mice in the Huanglian Jiedutang group and the Ginaton group were administered the corresponding drugs by oral gavage once daily at a volume of 0.15 mL·（10 g）^-1 for 7 consecutive days， while the sham-operated and model groups received an equal volume of saline via the same route. After 7 days， MCAO surgery was performed. The distal and proximal ends of the right common carotid artery （CCA） were ligated， a small incision was made between the two ligatures， and a silicone rubber-coated monofilament with a rounded tip was inserted into the lumen to occlude the CCA. The filament was left in place for 1 h to establish a focal cerebral ischemia model. At 24 h after modeling， mice were evaluated. Neurological function was assessed using the Longa score. Cerebral infarct volume was measured by 2，3，5-triphenyltetrazolium chloride （TTC） staining. Cerebral blood flow was observed by laser speckle imaging. Hematoxylin and eosin （HE） staining and Nissl staining were used to observe pathological changes in brain tissues. Exosomes were isolated from mouse plasma and brain tissues by ultracentrifugation and molecular size exclusion and identified by electron microscopy， particle size analysis， and protein blotting. Long-chain RNA libraries of exosomes were constructed and sequenced. Real-time quantitative reverse transcription polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of inflammatory factors and neutrophil-related chemokines in exosomes from plasma and brain tissues of each group. Enzyme-linked immunosorbent assay （ELISA） was used to detect the protein expression of inflammatory factors and neutrophil-related chemokines in exosomes from brain tissues of each group. Immunohistochemistry was used to detect the expression of the neutrophil-specific protein myeloperoxidase （MPO） in the brains of mice in each group. ResultsCompared with the sham-operated group， the model group showed decreased neurological function scores （P<0.01）， obvious cerebral infarction （P<0.01）， reduced cerebral blood flow （P<0.01）， neuronal necrosis in the brain， and decreased numbers of Nissl bodies （P<0.01）. The mRNA expression levels of IL-1β， MPO， CXCL1， CXCL2， CXCL3， CXCL10， CCL2， and CCL3 in exosomes from plasma and brain tissues were significantly increased （P<0.05， P<0.01）. The protein expression levels of IL-1β， MPO， CXCL2， and CXCL10 in exosomes from brain tissues were increased （P<0.05， P<0.01）， and MPO-positive rates and mean optical density values in brain tissues were elevated （P<0.01）. Compared with the model group， the Huanglian Jiedutang group and the Ginaton group showed increased neurological function scores （P<0.05）， reduced cerebral infarct volume （P<0.01）， restored cerebral blood flow （P<0.01）， reduced necrotic cells in the brain， and increased numbers of Nissl bodies （P<0.01）. In the Huanglian Jiedutang group， the mRNA expression levels of IL-1β， MPO， CXCL1， CXCL2， CXCL3， CXCL10， CCL2， and CCL3 in exosomes from plasma and brain tissues were decreased （P<0.05， P<0.01）. The protein expression levels of IL-1β， MPO， CXCL2， and CXCL10 in exosomes from brain tissues were reduced （P<0.05， P<0.01）， and MPO-positive rates and mean optical density values in brain tissues were decreased （P<0.01）. ConclusionHuanglian Jiedutang can effectively regulate the expression of neutrophil-related chemokines in exosomes from plasma and brain tissues of MCAO mice， thereby reducing neutrophil infiltration in the brain and achieving therapeutic effects.

ObjectiveTo investigate the effect and molecular mechanism of exosomes derived from bone marrow mesenchymal stem cells（BMSC-exos） modified with Bushen Yisui capsule（BSYS）-containing serum on promoting the differentiation and maturation of OLN-93 oligodendrocytes by regulating miR-15b/Wnt signaling pathway. MethodsOLN-93 cells were divided into 5 groups， including the normal（NC） group， BMSC-exos group， BSYS-BMSC-exos group， BSYS-BMSC+LV-miR-15b-5p inhibitor-exos group， and BSYS-BMSC+LV-miR-15b-5p NC-exos group. DiR staining was used to observe the uptake of Exos by OLN-93 cells. The effective dosage of BSYS-BMSC-exos on OLN-93 cells was assessed by cell proliferation and activity assay（CCK-8）. Stable BMSCs lentiviral transfection strains were established to inhibit miR-15b-5p expression in both BMSCs and their exos， and transfection efficiency was verified by real-time fluorescent quantitative polymerase chain reaction（Real-time PCR） detection of miR-15b-5p. The expressions of 2′，3′-cyclic nucleotide 3′-phosphodiesterase（CNPase） and myelin proteolipid protein（PLP） in OLN-93 cells were detected by immunocytochemistry（ICC） and Western blot. The mRNA expressions of miR-15b-5p and Wnt3a in OLN-93 cells were detected by Real-time PCR， and the protein expression of Wnt3a was measured by Western blot. The expression levels of key molecules in the Wnt/β-catenin signaling pathway of OLN-93 cells， including glycogen synthase kinase（GSK）-3β， β-catenin， and T-cell specific transcription factor 4/transcription factor 7-like 2（TCF4/TCF7L2）， were measured by Real-time PCR and Western blot. ResultsDiR-labeled Exos were efficiently taken up by OLN-93 cells. The CCK-8 assay results indicated that 20 mg·L^-1 of BSYS-BMSC-exos exhibited the most significant effect in enhancing OLN-93 cell viability（P<0.01） and this dosage was selected for subsequent experiments. Following lentiviral transfection of BMSCs， Real-time PCR results revealed that miR-15b-5p was significantly suppressed in BMSCs（P<0.01）， and miR-15b-5p was also notably inhibited in BSYS-BMSC-exos（P<0.01）. ICC analysis further revealed an increase in the number of differentiated， mature CNPase and PLP-positive cells following BSYS-BMSC-exos treatment（P<0.01）. Western blot results demonstrated that the protein expression of CNPase and PLP was significantly enhanced with BSYS-BMSC-exos treatment（P<0.01）. Additionally， BSYS-BMSC-exos also increased the expression levels of miR-15b-5p and p-β-catenin proteins in OLN-93 cells， while decreased the mRNA and protein expressions of Wnt3a， as well as the mRNA expressions of β-catenin and TCF4/TCF7L2， and the protein expression level of p-GSK-3β（Ser9） was significantly reduced（P<0.05， P<0.01）. After the transfection of miR-15b-5p inhibitor into BSYS-BMSC-exos， the above effects were significantly diminished（P<0.05， P<0.01）. ConclusionBSYS-BMSC-exos facilitate the differentiation and maturation of OLN-93 cells， and its mechanism is related to the upregulation of miR-15b-5p in OLN-93 cells， which inhibits the expression of Wnt3a and thereby suppresses the Wnt signaling pathway.

Objective To investigate the bone protective effects and underlying mechanisms of 900MHz radiofrequency radiation (RF) at different power densities (50, 150, and 450 μW/cm²) on an ovariectomy-induced osteoporosis mouse model. Methods Sixty 3-month-old C57BL/6 female mice were randomly divided into Sham group (sham exposure), OVX group (ovariectomy), OVX + LRF group (OVX + 50 μW/cm² RF), OVX + MRF group (OVX + 150 μW/cm² RF), OVX + HRF group (OVX + 450 μW/cm² RF), and OVX + E2 group (OVX + estradiol). Ovariectomized mice in the OVX + RF groups were exposed to RF of varying power densities for 4 hours daily. Ovariectomized mice in the OVX + E2 group received intramuscular injections of estradiol (0.04 mg/kg) every two days. After four weeks of intervention, Micro-CT, ELISA, RT-PCR, and immunohistochemistry were employed to analyze bone density, bone microstructure, serum bone metabolic markers, and the expression of related genes and proteins. Results Compared with the Sham group, the OVX group showed significantly decreased bone mineral density (BMD) and bone microstructure indicators such as BV, BV/TV, Tb.Th, and Tb.N, significantly increased bone microstructure indicator Tb.Sp, significantly decreased serum estradiol, significantly increased serum CTX-I, TRACP-5b, BGP, and OPG, significantly increased Nfatc1 and Runx2 mRNAs, and significantly increased OPG and RANKL. Compared with the OVX group, the OVX + MRF group and OVX + E2 group exhibited significantly increased BMD, BV, BV/TV, Tb.Th, and Tb.N, significantly decreased Tb.Sp, significantly increased serum OPG, Runx2 mRNA, and OPG, and significantly decreased serum CTX-I, TRACP-5b, Nfatc1 mRNA, and RANKL. Compared with the OVX group, the OVX + LRF group showed significantly increased cortical bone BMD and Tb.Th, the OVX + HRF group showed significantly increased cortical bone BMD and serum CTX-I and TRACP-5b, and the OVX + MRF group showed significantly increased serum BGP. Among the three power densities, the 150 μW/cm² RF showed the most significant effect. Conclusion The 150 μw/cm² 900 MHz RF can counteract the abnormalities in serum bone metabolism biomarkers, the decrease in BMD, the degeneration of bone microstructure, and the increase in bone resorption caused by ovariectomy in mice.

To investigate the association between neutrophil to lymphocyte ratio (NLR) andestimated glomerular filtration rate (eGFR) in patients with diabetes using large-scale data.

Objective To explore the influence of the radiochemotherapy mode and the number of syn-chronized chemotherapy cycles on the efficacy and overall survival(OS)time of the patients with limited-stage small cell lung cancer(LS-SCLC).Methods The clinical data of 85 patients with LS-SCLC receiving the thoracic radiotherapy(TRT)in the tumor radiotherapy department of this hospital from January 2018 to De-cember 2022 were retrospectively analyzed.The patients were divided into the synchronous chemotherapy group(n=43)and non-synchronous chemotherapy group(n=42)according to different radiochemotherapy modes.The OS and progress free survival(PFS)time was compared between the two groups,and the related prognostic influencing factors of LS-SCLC were analyzed.Results The last follow-up date was November 1,2023,and the median follow-up time in all groups was 39.6 months.The median OS time of the synchronous chemotherapy group and non-synchronous chemotherapy group was 30.6 months and 18.6 months respective-ly,and the difference was statistically significant(P<0.05).The median OS time of 1 cycle and 2 cycles in the synchronous chemotherapy group was 25.5 months and 33.6 months,and the difference was not statisti-cally significant(P>0.05).The Cox multivariate regression analysis results showed that the age,clinical stage and prophylactic cranial irradiation(PCI)were the independent prognostic influencing factors in the pa-tients with LS-SCLC.Conclusion The number of synchronized chemotherapy cycles has no significant influ-ence on the prognosis of LS-SCLC.

Objective:To explore the clinical efficacy of acupuncture combined with hemoperfusion in hemodialysis patients with intractable pruritus.Methods:A retrospective clinical study was conducted. A total of 126 hemodialysis patients with intractable pruritus were enrolled from June 2023 to June 2024 and divided into two groups (63 cases each) based on treatment modalities. The control group received hemoperfusion alone, while the observation group received acupuncture combined with hemoperfusion. Both groups were treated for 3 months. The Modified Pruritus Duo's Score was used to evaluate pruritus severity, the Pittsburgh Sleep Quality Index (PSQI) for sleep quality, and the 36-Item Short Form Health Survey (SF-36) for quality of life before and after treatment. Serum β 2-microglobulin (β 2-MG) and parathyroid hormone (PTH) levels were measured using immunoturbidimetry and chemiluminescence, respectively. Clinical efficacy was assessed. Results:The total effective rate was 98.42% (62/63) in the observation group and 78.77% (49/63) in the control group, with a statistically significant difference ( χ2=16.90, P<0.001). Post-treatment scores for pruritus severity, affected areas, frequency, sleep interference, and total pruritus score in the observation group were significantly lower than those in the control group ( t values were 16.94, 12.23, 12.67, 12.71, 23.91, respectively, P<0.001). The observation group also exhibited lower post-treatment serum β 2-MG [(13.24±3.11) mg/L vs. (17.61±4.58) mg/L, t=10.46] and PTH [(198.11±96.12) ng/L vs. (249.27±88.45) ng/L, t=6.72] levels were lower than those in the control group ( P<0.001). Additionally, the observation group demonstrated significantly lower scores in subjective sleep quality, sleep latency, sleep duration, habitual sleep efficiency, sleep disturbances, daytime dysfunction, and total PSQI score ( t values were 14.97, 6.98, 2.06, 23.63, 2.68, 3.56, 14.89, respectively, P<0.001), as well as higher scores in physical health, role-physical, bodily pain, general health, vitality, social functioning, role-emotional, and mental health domains of SF-36 ( t values were -3.13, -4.35, -6.34, -7.90, -4.49, -5.30, -5.00, -3.95, respectively, P<0.05). Conclusion:Acupuncture combined with hemoperfusion effectively alleviates pruritus, reduces serum β 2-MG and PTH levels, improves sleep quality, and enhances overall quality of life in hemodialysis patients with intractable pruritus, demonstrating superior efficacy compared to hemoperfusion alone.

Protein arginine methyltransferase 5 (PRMT5) acts as an oncogene in liver cancer, yet its roles and in-depth molecular mechanisms within the liver cancer immune microenvironment remain mostly undefined. Here, we demonstrated that disruption of tumor-intrinsic PRMT5 enhances CD8⁺ T-cell-mediated antitumor immunity both in vivo and in vitro. Further experiments verified that this effect is achieved through downregulation of the inhibitory immune checkpoint molecule, fibrinogen-like protein 1 (FGL1). Mechanistically, PRMT5 catalyzed symmetric dimethylation of transcription factor 12 (TCF12) at arginine 554 (R554), prompting the binding of TCF12 to FGL1 promoter region, which transcriptionally activated FGL1 in tumor cells. Methylation deficiency at TCF12-R554 residue downregulated FGL1 expression, which promoted CD8⁺ T-cell-mediated antitumor immunity. Notably, combining the PRMT5 methyltransferase inhibitor GSK591 with PD-L1 blockade efficiently inhibited liver cancer growth and improved overall survival in mice. Collectively, our findings reveal the immunosuppressive role and mechanism of PRMT5 in liver cancer and highlight that targeting PRMT5 could boost checkpoint immunotherapy efficacy.