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.

OBJECTIVE To investigate the effects of galangin on rheumatoid arthritis （RA） in rats by regulating the Janus kinase 3 （JAK3）/signal transducer and activator of transcription 3 （STAT3） pathway. METHODS Fifty male SD rats were taken， and an emulsion composed of bovine type Ⅱ collagen and Freund’s complete adjuvant was injected subcutaneously to establish an induced arthritis model. The rats that were successfully modeled were randomly divided into model group， low， medium and high dose groups of galangin （1， 5， 15 mg/kg）， and methotrexate group （positive control， 2 mg/kg）， with 10 rats in each group. Another 10 normal rats were taken as the normal group. Starting from the 15th day of modeling， each group of rats was gavaged with the corresponding drug solution or normal saline containing 0.5% Tween 80 once a day for 28 consecutive days. The arthritis index （AI） scores and paw volume of rats were compared before and after gavage administration. Twenty-four hours after the last administration， the serum levels of interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， IL-4 and IL-10 were determined， the pathological changes in ankle joint synovial tissue were observed， and the protein expressions of UNC-51 like kinase 1 （ULK1）， Beclin-1， microtubule-associated protein 1 light chain 3 （LC3）， B cell lymphoma 2 （Bcl-2）， Bcl-2-associated X protein （Bax）， caspase-3， JAK3， phosphorylated JAK3 （p-JAK3）， STAT3 and phosphorylated STAT3 （p-STAT3） in the synovial tissue of the ankle joint were detected， as well as the fluorescence intensity of LC3-positive areas. RESULTS Compared with the model group， the pathological changes such as cellular proliferation of ankle joint synovial tissue and infiltration of inflammatory cells in rats of each administration group showed improvement. Moreover， their AI scores and paw pad volumes （on day 28 after gavage）， the levels of IL-6 and TNF-α， the protein expression of Bcl-2， and the phosphorylation levels of JAK3 and STAT3 were all significantly reduced （ P ＜0.05）. The levels of IL-4 and IL-10， the protein expressions of ULK1， Beclin-1， Bax， caspase-3 and LC3， as well as the fluorescence intensity of LC3-positive areas， were all significantly increased （ P ＜0.05）. Moreover， the effect of galangin was in a dose-dependent manner （ P ＜0.05）. CONCLUSIONS Galangin can induce sustained autophagy in synovial tissue cells of RA rats， promote cell apoptosis， inhibit synovial cell proliferation， and alleviate persistent inflammatory responses. The above anti-RA effects may be related to the inhibition of the JAK3/STAT3 pathway.

Circulating tumor cells (CTCs) have the ability to sow tumors and can be found in the peripheral blood of patients with precancerous lesions and healthy people. However, CTCs are not currently screened in the donors blood. A large number of allogeneic blood transfusions occurred worldwide each year, and allogeneic blood transfusions expose recipients to the risk of transmission and affect tumors associated with donor CTCs. Although leukocyte filtration can not completely remove tumor cells in the blood, it can effectively reduce the number of white blood cells in the blood and reduce their proliferation ability. Blood irradiation can effectively destroy the DNA of CTCs in the blood, and inhibit the occurrence and metastasis of tumors caused by the infusion of allogeneic blood containing CTCs. Therefore, we should pay attention to the potential risk of CTCs on clinical transfusion, and strengthen the preclinical treatment of blood to avoid donor-related tumor infection in blood recipients due to clinical transfusion.

ObjectiveTo explore the current situation and influencing factors of humanistic care experience among patients visiting traditional Chinese medicine （TCM） outpatient clinics in China， and to provide a basis for optimizing TCM-characterized services in both TCM and Western medicine hospitals. MethodsA multi-center cross-sectional study was conducted using convenience sampling to select 35 hospitals across 13 provinces in China （including 3 TCM hospitals and 32 TCM outpatient clinics in general hospitals）. A total of 3，430 patients were surveyed using the general information questionnaire and the Outpatient Humanistic Care Experience Questionnaire， with data collected via Questionnaire Star. Univariate analysis and multiple linear regression were employed to examine the impacts of patient characteristics， visit characteristics， hospital type （TCM hospital/general hospital）， and geographic region （eastern/central/western） on humanistic care experience. ResultsThe total score of humanistic care experience was 194 （188， 233）. Univariate analysis showed that gender， educational level， current residence， per capita monthly household income， location attribute of medical institutions， number of previous visits to this hospital， payment method of medical expenses， previous hospitalization history in this hospital， frequency of outpatient visits within the past 12 months， self-rated disease severity， familiarity with the outpatient procedures， implementation of the follow-up service provided by the hospital， satisfaction with follow-up services， the grade of the hospital visited， geographical region of the hospital visited， and the department visited had an impact on the humanistic care experience during outpatient visits （P<0.05）. Multivariate analysis demonstrated that educational level （β=0.609， P=0.011）， self-rated disease severity （β=-0.646， P=0.047）， familiarity with outpatient procedures （β=4.784， P<0.001）， satisfaction with follow-up services （β=6.365， P<0.001）， and the grade of the hospital visited （β=-5.487， P<0.001） affected the humanistic care experience in outpatient medical treatment， explaining 24.4% of the total variation. ConclusionHumanistic care experience in TCM outpatient clinics is influenced by multiple factors. It is recommended to optimize the medical treatment process， strengthen doctor-patient communication training， and establish a precise follow-up mechanism， with a focus on improving care perceptions among patients with lower education levels and those attending primary-level hospitals， to refine the TCM-characterized service system.

BACKGROUND: Regulatory dendritic cell (DCreg) subset exhibits a unique capacity for inducing immune tolerance among the variety subsets of dendritic cells (DCs) within gut-associated lymphoid tissues (GALTs). Fms-like tyrosine kinase 3 ligand (FLT3L) is involved in the differentiation of DCregs and the subsequent expansion of regulatory T-cells (Tregs) mediated by DCregs, though the precise mechanism remains poorly understood. This study aimed to explore the expansion mechanism of Treg induced by DCreg and the role of FLT3L in this process. METHODS: DCregs were distinguished from other DC subsets isolated from GALTs of BALB/c mice through a mixed lymphocyte reaction assay. The functions and mechanisms by which FLT3L promoted Treg expansion via DCregs were investigated in vitro through co-culture experiments involving DCregs and either CD4 + CD25 - T-cells or CD4 + CD25 + T-cells. Additionally, an in vivo experiment was conducted using a dextran sulfate sodium (DSS)-induced colitis model in mice. RESULTS: CD103 + CD11b + DC exhibited DCreg-like functionality and was identified as DCreg for subsequent investigation. Analysis of Foxp3 + Treg percentages within a co-culture system of CD4 + CD25 - T-cells and DCregs, with or without FLT3L, demonstrated the involvement of the FLT3/FLT3L axis in driving the differentiation of precursor T-cells into Foxp3 + Tregs induced by DCregs. Cell migration and co-culture assays revealed that the FLT3/FLT3L axis enhanced DCreg migration toward Tregs via the Rho pathway. Additionally, it was observed that DCregs could promote Treg proliferation through the Notch pathway, as inhibition of Notch signaling by DAPT (N-[N-(3,5-difluorophenacetyl)-l-alanyl]-S-phenylglycine t-butyl ester) suppressed Treg expansion within the co-culture system of DCregs and CD4 + T-cells or CD4 + CD25 + T-cells. Furthermore, the FLT3/FLT3L axis influenced JAG1 expression in DCregs, indirectly modulating Treg expansion. In vivo experiments further established that FLT3L promoted DCreg expansion and restored Treg balance in DSS-induced colitis models, thereby ameliorating colitis symptoms in mice. CONCLUSION The FLT3/FLT3L axis is integral to the maintenance of DCreg function in Treg expansion.
Animals ; T-Lymphocytes, Regulatory/immunology* ; Dendritic Cells/immunology* ; Mice ; Mice, Inbred BALB C ; Membrane Proteins/metabolism* ; Receptors, Notch/metabolism* ; Lymphoid Tissue/metabolism* ; Signal Transduction/physiology* ; Coculture Techniques ; Flow Cytometry

Coronavirus disease 2019 (COVID-19) is a multi-system disease that can lead to various severe complications. Acute limb ischemia (ALI) has been increasingly recognized as a COVID-19-associated complication that often predicts a poor prognosis. However, the pathophysiology and molecular mechanisms underlying COVID-19-associated ALI remain poorly understood. Hypercoagulability and thrombosis are considered important mechanisms, but we also emphasize the roles of vasospasm, hypoxia, and acidosis in the pathogenesis of the disease. The angiotensin-converting enzyme 2 (ACE2) pathway, inflammation, and platelet activation may be important molecular mechanisms underlying these pathological changes induced by COVID-19. Furthermore, we discuss the hypotheses of risk factors for COVID-19-associated ALI from genetic, age, and gender perspectives based on our analysis of molecular mechanisms. Additionally, we summarize therapeutic approaches such as use of the interleukin-6 (IL-6) blocker tocilizumab, calcium channel blockers, and angiotensin-converting enzyme inhibitors, providing insights for the future treatment of coronavirus-associated limb ischemic diseases.
Humans ; COVID-19/physiopathology* ; Ischemia/etiology* ; SARS-CoV-2 ; Extremities/blood supply* ; Risk Factors ; Interleukin-6/antagonists & inhibitors* ; Acute Disease ; Angiotensin-Converting Enzyme 2

Normal heart function depends on complex regulation by the brain, and abnormalities in the brain‒heart axis affect various diseases, such as myocardial infarction and anxiety disorders. However, systematic tracking of the brain regions associated with the input and output of the heart is lacking. In this study, we injected retrograde transsynaptic pseudorabies virus (PRV) and anterograde transsynaptic herpes simplex virus (HSV) into the left ventricular wall of mice to identify the whole-brain regions associated with the input to and output from the heart. We successfully detected PRV and HSV expression in at least 170 brain subregions in both male and female mice. Sex differences were discovered mainly in the hypothalamus and medulla, with male mice exhibiting greater correlation and hierarchical clustering than female mice, indicating reduced similarity and increased modularity of virus expression patterns in male mice. Further graph theory and multiple linear regression analysis of different injection timelines revealed that hub regions of PRV had highly similar clusters, with different brain levels, suggesting a top-down, hierarchically transmitted neural control pattern of the heart. Hub regions of HSV had scattered clusters, with brain regions gathered in the cortex and brainstem, suggesting a bottom-up, leapfrog, multipoint neural sensing pattern of the heart. Both patterns contain many hub brain regions that have been previously overlooked in brain‒heart axis studies. These results provide brain targets for future research and will lead to deeper insight into the brain mechanisms involved in specific heart conditions.
Animals ; Male ; Female ; Heart/physiology* ; Mice ; Herpesvirus 1, Suid ; Brain/physiology* ; Mice, Inbred C57BL ; Brain Mapping ; Efferent Pathways/physiology* ; Afferent Pathways/physiology* ; Simplexvirus ; Sex Characteristics

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.

Objective: To investigate the prevalence and influencing factors of menopausal syndrome among postmenopausal women in Pan'an County, Zhejiang Province, so as to provide the basis for guiding the health management of postmenopausal women. Methods: From May 2023 to April 2024, the postmenopausal women aged 40 to 69 years in Pan'an County were selected using the random cluster sampling method. Demographic information, lifestyle and prevalence of gynecological diseases were collected through questionnaire surveys. The prevalence of menopausal syndrome was assessed by modified Kupperman Score Scale. Factors affecting menopausal syndrome were analyzed by a multivariable logistic regression model. Results: A total of 816 postmenopausal women were surveyed, with an mean age of (57.63±2.92) years and a mean natural menopause age of (49.85±2.13) years. There were 574 cases with menopausal syndrome, with a prevalence of 70.34%. Flashes and sweating, insomnia and irritability were common symptoms, accounting for 62.87%, 47.43% and 41.18%, respectively. Multivariable logistic regression analysis showed that monthly personal income of ≤5 000 yuan (<3 000 yuan, OR=3.124, 95%CI: 1.829-5.335; 3 000-5 000 yuan, OR=2.399, 95%CI: 1.370-4.201) and having gynecological diseases (OR=1.970, 95%CI: 1.292-3.004) were associated with a higher risk of menopausal syndrome, while average (OR=0.141, 95%CI: 0.072-0.276) or sufficient sleep quality (OR=0.095, 95%CI: 0.049-0.185) were associated with a lower risk of menopausal syndrome. Conclusion The prevalence of menopausal syndrome among postmenopausal women in Pan'an County is relatively high, and is mainly influenced by personal economic status, sleep quality and the presence of gynecological diseases.