BACKGROUND:Eucommia ulmoides has a certain osteogenic effect,which can promote the proliferation and differentiation of osteoblasts.However,it is unclear whether Eucommia ulmoides has effects on alveolar bone formation and Wnt/β-Catenin signaling pathway. OBJECTIVE:To investigate the mechanism by which Eucommia ulmoides promotes alveolar bone formation in ovariectomized rats based on the Wnt/β-Catenin signaling pathway. METHODS:Sixty female Sprague-Dawley rats were selected and randomly divided into five groups:blank control group,sham-operation group,model group,low-dose group Eucommia ulmoides group,and high-dose Eucommia ulmoides group,with twelve rats in each group.Osteoporosis animal models were constructed by bilateral oophorectomy in the model group and the low-dose and high-dose Eucommia ulmoides groups.The sham-operation group underwent the same method to remove adipose tissue of equal mass around the bilateral ovaries.Three months after surgery,the low-and high-dose Eucommia ulmoides groups were given 2.1 g/kg/d and 4.2 g/kg/d Eucommia ulmoides by gavage,respectively.The sham-operation group and model group were given the same amount of physiological saline by gavage.After 12 weeks of drug intervention,the changes in alveolar bone mass of rats in each group were observed through Micro-CT;hematoxylin-eosin staining was used to observe the pathological structural changes of alveolar bone in rats;enzyme linked immunosorbent assay was used to detect the expression levels of alkaline phosphatase and osteocalcin in the serum of rats;western blot was used to detect the expression levels of β-Catenin and Frizzled9 receptor proteins in the alveolar bone of rats;and real-time fluorescence quantitative PCR was used to detect the expression of osteocalcin,Runt-related transcription factor 2(Runx2),alkaline phosphatase,β-catenin,and frizzled9 mRNAs in alveolar bone tissues of rats. RESULTS AND CONCLUSION:Compared with the blank control group,bone volume fraction,trabecular number,trabecular thickness,and bone mineral density were reduced in the model group(P＜0.05),and trabecular separation was elevated(P＜0.05).Pathological observation showed that the arrangement of trabeculae was disordered and irregular,the trabeculae were thinned or broken,and the marrow cavity was enlarged in the model group,with a significant reduction in bone volume;the level of alkaline phosphatase in the serum was increased(P＜0.05),and the level of osteocalcin was decreased(P＜0.05);mRNA expression of alkaline phosphatase,osteocalcin,Runx2,β-catenin,and frizzled9 were decreased(P＜0.05);protein expression of β-Catenin and Frizzled9 was decreased(P＜0.05).Compared with the model group,the low-and high-dose Eucommia ulmoides groups showed an increase in bone volume fraction,trabecular number,trabecular thickness,and bone mineral density(P＜0.05)and a decrease in trabecular separation(P＜0.05).In the low-and high-dose Eucommia ulmoides groups,bone trabeculae were slightly aligned and thickened,with a significant increase in bone mass.Compared with the model group,the serum level of alkaline phosphatase was reduced(P＜0.05)and the serum level of osteocalcin was elevated(P＜0.05)in the low-and high-dose Eucommia ulmoides groups.Compared with the model group,the mRNA expression of alkaline phosphatase,osteocalcin,Runx2,β-catenin,and frizzled9 were increased in the low-and high-dose Eucommia ulmoides groups(P＜0.05).Compared with the model group,the protein expression of Frizzled9 was increased in the low-dose Eucommia ulmoides group(P＜0.05),while the protein expression of β-Catenin and Frizzled9 was increased in the high-dose Eucommia ulmoides group(P＜0.05).Compared with the low-dose Eucommia ulmoides group,the high-dose Eucommia ulmoides group had a more significant improvement in the above indexes.To conclude,Eucommia ulmoides can effectively promote the alveolar bone formation,and its mechanism of action might be related to the activation of the Wnt/β-catenin signaling pathway.

BACKGROUND: Cluster of differentiation 8 positive (CD8 + ) T cells play a crucial role in the response against tumors, including hepatocellular carcinoma (HCC), where their dysfunction is commonly observed. While the association between elevated peroxisome proliferator-activated receptor alpha (PPARα) expression in HCC cells and exosomes and unfavorable prognosis in HCC patients is well-established, the underlying biological mechanisms by which PPARα induces CD8 + T cell exhaustion mediated by HCC exosomes remain poorly understood. METHODS: Bioinformatics analyses and dual-luciferase reporter assays were used to investigate the regulation of microRNA-27b-3p ( miR-27b-3p ) and thymocyte selection-associated high mobility group box ( Tox ) by Pparα . In vitro and in vivo experiments were conducted to validate the effects of HCC-derived exosomes, miR-27b-3p overexpression, and Pparα on T cell function. Exosome characterization was confirmed using transmission electron microscopy, Western blotting, and particle size analysis. Exosome tracing was performed using small animal in vivo imaging and confocal microscopy. The expression levels of miR-27b-3p , Pparα , and T cell exhaustion-related molecules ( Tox , Havcr2 , and Pdcd1 ) were detected using quantitative reverse transcription polymerase chain reaction analysis, Western blotting analysis, immunofluorescence staining, and flow cytometry analysis. RESULTS: Pparα expression was significantly increased in HCC and negatively correlated with prognosis. It showed a positive correlation with Tox and a negative correlation with miR-27b-3p . The overexpressed Pparα from HCC cells was delivered to CD8 + T cells via exosomes, which absorbed miR-27b-3p both in vitro and in vivo , acting as "miRNA sponges". Further experiments demonstrated that Pparα can inhibit the negative regulation of Tox mediated by miR-27b-3p through binding to its 3'untranslated regions. CONCLUSIONS HCC-derived exosomes deliver Pparα to T cells and promote CD8 + T cell exhaustion and malignant progression of HCC via the miR-27b-3p /TOX regulatory axis. The mechanisms underlying T-cell exhaustion in HCC can be utilized for the advancement of anticancer therapies.
MicroRNAs/metabolism* ; PPAR alpha/genetics* ; Carcinoma, Hepatocellular/genetics* ; Humans ; Liver Neoplasms/genetics* ; CD8-Positive T-Lymphocytes/immunology* ; Exosomes/metabolism* ; Animals ; Cell Line, Tumor ; Mice ; High Mobility Group Proteins/genetics* ; Male ; T-Cell Exhaustion

Objective To explore the effect of metformin combined with insulin detemir on intestinal flora in patients with gestational diabetes mellitus (GDM). Methods A total of 176 patients with GDM admitted to Luzhou People's Hospital from May 2022 to July 2023 were selected and randomly divided into a single drug group and a combined group, with 88 cases in each group. The single drug group was treated with insulin detemir, and the combined group was given metformin combined with insulin detemir. The glucose metabolism levels and intestinal flora distribution were compared between the two groups before treatment and during delivery. The maternal-infant outcomes were statistically analyzed and compared between the two groups. Results During delivery, the levels of fasting plasma glucose (FPG), glycosylated hemoglobin (HbAlc), and largest amplitude of glycemic excursions (LAGE), and the contents of Enterobacterium, Enterococcus and Escherichia coli in both groups were reduced compared with those before treatment, and the above indicators in the combined group were lower than those in the single drug group (all P<0.05). The contents of Bifidobacterium and Lactobacillus were increased in both groups, and the indicators were higher in the combined group than those in the single drug group (all P<0.05). There was no significant difference in adverse maternal-infant outcomes between the two groups (P>0.05). Conclusion Metformin combined with insulin detemir can effectively reduce blood glucose levels and improve intestinal flora distribution in patients with GDM, without increasing adverse maternal-infant outcomes.

Objective:To explore the associations of temperature, relative humidity, and their interaction effect with fall risk.Methods:Data on fall cases were collected using the national injury surveillance system from May to September, in 2006-2022 in Hubei Province. Combined with the meteorological and air pollution data, we conducted a time-stratified case-crossover design and used conditional logistic regression models and distributed lag nonlinear models to examine the exposure-response relationships between temperature, humidity, and fall risk. We further divided the low and the high temperature groups and the low and the high relative humidity groups and analyzed the excess risk ( ER) of falls attributed to dry-hot or wet-hot events. Finally, we calculated the additive interactions of temperature and humidity on fall risk. Results:A total of 55 401 fall cases were included. With the increase in temperature and relative humidity decrease, the exposure-response curves of fall showed nonlinear upward trends among all populations. Gender and age differences were found in temperature-fall and relative humidity-fall risk relationships. Compared with wet-non-hot (normal temperature and high relative humidity) events, the ER of fall in dry-hot (high temperature and low relative humidity) events was 14.80% (95% CI: 9.69%- 20.15%), and the ER of wet-hot (high temperature and high relative humidity) events was 9.59% (95% CI: 2.52%-17.13%). However, there was no statistically significant difference between dry-hot and wet-hot events in the fall, and no statistically significant difference between different genders, ages, occupations, and fall occurred place (all P>0.05). No significant synergistic additive interaction was found between temperature and relative humidity on fall risk (relative excess risk due to interaction=-0.08, 95% CI: -0.19-0.02). Conclusions:Higher temperatures and lower relative humidity were associated with increased fall risk. Both dry-hot and wet-hot events had a higher risk of fall, while high temperature and low humidity have no synergistic effect on fall risk.

Objective:To establish a rapid and precise dose measurement method with EBT4 film and ensure its measurement accuracy to be within the required range through strict operational procedures for the purpose of addressing the two essential issues of poor measurement accuracy and timeliness of EBT film under FLASH conditions.Methods:After storing under different humidity conditions for a certain period of time, the film was exposed to radiation for analyzing the influence of air humidity on the intrinsic performance of EBT film. By means of repeated scanning operations and the film angle rotation, the influences of repeated scanning and film placement angle were analuzed. Parabolic correction method was used to reduce the spatial position influence during the scanning process. By analying the relationship between net optical density (netOD) and absorbed dose through the comparison of three fitting method, the optimal fitting curve was selected. After irradiation of the same batch of films for 5 min and 24 h, the film doses were calibrated and then compared with ionization chamber-measured result. The rapid and precise film dosimetry method was used to measure both the percentage depth dose from X-rays at ultra-high dose rate and the dose distribution at a depth of 2 cm in water.Results:Air humidity had the greatest influence on the intrinsic performance of EBT film (approximately 20%). The average deviation of repeated scans is within 0.5%. The angle at which the film is placed significantly affected the readouts of the film with the maximum influence approximately 70%. The net optical density combined with polynomial fitting can control the fitting residuals of 1-16 Gy within 3%. The change rate of light channels at 5 min already mostly met the requirements of the rapid mode (< 0.5%). Compared with the measurement result obtained using the reference ionization chamber, the deviations of the 5 min or 24 h dose calibration curves were all within 2%.Conclusions:The EBT4 film can be employed as a precise dosimeter to quickly measure the FLASH radiation dose. Rapid and precise FLASH dose measurements can meet the stringent requirements of both preclinical and clinical FLASH research.

Objective:To investigate the feasibility and clinical value of artificial intelligence-assisted compressed sensing (ACS) technology in accelerating MR simulation (MR-sim) for radiotherapy of nasopharyngeal carcinoma (NPC).Methods:Thirty patients with NPC scheduled to receive radical radiotherapy at Sun Yat-sen University Cancer Center were prospectively enrolled. All patients underwent head and neck MR-sim on a 3.0 T scanner, with axial T 1 weighted imaging (WI), T 2WI, contrast-enhanced T 1WI, and fat-suppressed contrast-enhanced T 1WI images acquired using both ACS and parallel imaging (PI) techniques. Paired-sample t tests or rank-sum tests were used to compare scan time, signal-to-noise ratio (SNR), and contrast-to-noise ratio (CNR) of MR-sim images between the two techniques. A 5-point Likert scale was applied to evaluate tumor lesion visualization, lesion margin clarity, artifacts, and overall image quality, with chi-square tests used to compare subjective image quality scores between the two techniques. Tumor target volumes were delineated on MR-sim images obtained by both ACS and PI techniques after fusion with CT simulation images, and consistency was assessed using the Dice similarity coefficient (DSC). Results:For both individual sequences and overall protocols, ACS significantly reduced MR-sim acquisition time compared with PI ( P < 0.001). The total acquisition time with ACS was (378.60±17.07) s versus (694.93±17.07) s with PI, representing a 45.52% time reduction. SNR, CNR, tumor lesion identification, margin clarity, artifacts, and overall image quality scores of MR-sim images did not differ significantly between ACS and PI ( P > 0.05). Tumor target volumes delineated from ACS- and PI-based MR-sim images showed high consistency after fusion with CT simulation images ( P > 0.05), with mean DSC values of primary tumors and metastatic cervical lymph nodes approaching 1. Conclusion:Compared with conventional MR acceleration methods (PI), ACS enables faster MR-sim acquisition in NPC without compromising image quality or the accuracy of tumor target delineation.

Background::Intrauterine growth restriction (IUGR) is associated with adverse metabolic outcomes during adulthood. Histone modifications and changes in DNA methylation-affected genes are important for fetal development. This study aimed to investigate the epigenetic mechanisms in IUGR.Methods::IUGR models were established in Sprague–Dawley rats using a maternal nutritional restriction approach during pregnancy. The abundance of insulin-like growth factor 2 (IGF2), phosphoinositide 3-kinase (PI3K), AKT serine/threonine kinase 2 (AKT2), and peroxisome proliferators-activated receptor gamma coactivator 1 alpha (PGC-1α) was examined by real-time polymerase chain reaction (RT-PCR) and Western blotting analysis. Chromatin immunoprecipitation RT-PCR was employed to analyze histone modification in CCCTC-binding factor (CTCF) 1–4 binding sites of the Igf2/H19 imprinting control region (ICR). The methylation states of CTCF1–4 binding sites were studied by pyrosequencing. Results::The IUGR models were constructed successfully. Igf2 mRNA abundance in the placenta, fetal liver, and newborn liver was decreased in the IUGR group ( P <0.01). Meanwhile, as compared with the control group, the expression levels of AKT2, PI3K, and PGC-1α were lower in newborn and 8-week-old livers in the IUGR group ( P <0.05). In addition, knocking down Igf2 reduced the protein expression levels of AKT2-phosphorylation and PGC-1α ( P <0.05). In CTCF binding sites 1-4 of the Igf2/ H19 ICR, acetylated histones H3 (AcH3) enrichment was significantly lower in CTCF1-3 in newborn and 8-week-old IUGR rats. Histone H3 tri-methylated lysine 4 (H3K4me3) enrichment was significantly lower in the CTCF1–4 of newborn and 8-week-old IUGR groups ( P <0.01). H3K9me2 enrichment was significantly higher in the IUGR group ( P <0.01). The CpG dinucleotide methylation levels of CTCF1 and CTCF3, but not those of CTCF2 and CTCF4 binding sites in IUGR rat fetal, 4-week old, and 8-week-old livers decreased significantly ( P <0.05). Conclusion::The methylation status and histone modification in the Igf2/H19 ICR are related to growth and lipid metabolism via the PGC-1α/PI3K/AKT2 pathway in IUGR rats.

Objective In this study,we aimed to predict the inhibitory mechanism of Shenlingcao oral liquid(SLC)in non-small cell lung cancer(NSCLC)by network pharmacology and verify it by molecular docking and in vivo experiments.Methods The active ingredients and corresponding targets of SLC and NSCLC were obtained by database and literature search.Targets of SLC common to NSCLC were selected to construct the protein interaction network,and GO and KEGG enrichment analysis and molecular docking were performed.A Lewis lung cancer mouse model was constructed and divided into Model group,SH group,and SL group.The latter two groups were intragastrically administered 8.75 g SLC lyophilized powder/kg and 3.50 g SLC lyophilized powder/kg,respectively.After 14 days of drug intervention,tumor growth,pathological changes in tumor tissue,and apoptosis in tumor tissue were observed in tumor-bearing mice;changes in blood routine indexes and the tumor tissue expression of p-AKT,AKT,p-PI3K,PI3K,and Bcl-2 protein of mice were detected.The result of the KEGG enrichment analysis were verified.Results Network pharmacological analysis showed that there were 77 active ingredients,618 potential targets,1498 potential targets for NSCLC,and 179 drug and disease intersection targets.Target intersection enrichment analysis showed that they were mainly concentrated in the phosphatidylinositol 3 kinase-protein kinase B(PI3K-AKT)signaling pathway,mitogen-activated protein kinase(MAPK)signaling pathway,and other related pathways.Molecular docking showed that the top 10 core components had good bonding ability with the top 10 core targets.In the animal experiments,compared with the Model group,SH group and SL group had significantly decreased tumor volume and weight(P＜0.05,P＜0.01),and significantly decreased white blood cell,neutrophil,and monocyte numbers(P＜0.01,P＜0.001).Red blood cells,platelets,and hemoglobin were significantly increased(P＜0.05,P＜0.01,P＜0.001);apoptotic cells were significantly increased in early tumor tissue(P＜0.05,P＜0.01),and the protein expression levels of p-PI3K/PI3K,p-AKT/AKT,and Bcl-2/GAPDH were significantly decreased(P＜0.05,P＜0.01).The expression levels of PI3K,AKT1,and Bcl-2 genes were significantly decreased(P＜0.05,P＜0.01).Conclusions The mechanisms of SLC activity against NSCLC may be related to the activation of the PI3K-AKT pathway and the promotion of apoptosis.

Objective To explore the mediating effect of empowerment ability between type D personality and self-management behavior of patients with diabetes mellitus(DM).Methods A total of 738 patients with type 2 diabetes mellitus(T2DM)hospitalized in the Department of Endocrinology of three tertiary hospitals in Hainan Province from December 2022 to May 2023 were selected and divided into Type D personality(Type D,n=104)group and T2DM group(n=634).The general data,biochemical indexes,scores of negative emotion(NA),social inhibition(SI),empowerment ability,and scale of DM self-management activities(SDSCA)were compared between the two groups,and the correlation between type D personality,empowerment ability and self-management ability was analyzed.The mediating effect model was used to analyze the mediating effect of empowerment ability on the four self-management behaviors of patients with type D personality,and the chain mediating effect model was used to analyze the relationship between type D personality,empowerment ability,self-management behaviors and HbA1c.Results Compared with the T2DM group,HbA1c,proportion of rural residence,proportion of complications≥3,proportion of education level of junior high school or above,proportion of monthly income<3000 yuan,and NA and SI scores were significantly higher in the Type D group(P<0.05).The empowerment ability and scores of healthy diet,regular exercise,blood glucose monitoring and medication compliance were lower in the Type D group than in the T2DM group(P<0.05).Spearman correlation analysis showed that the empowerment ability score was positively correlated with the scores of healthy diet,regular exercise,blood glucose monitoring and medication compliance(P<0.05).NA and SI scores were negatively correlated with empowerment ability score,healthy diet,regular exercise,blood glucose monitoring and medication compliance(P<0.05).The results of model analysis with empowerment ability as the mediating variable showed that type D personality had direct,indirect and total effects on regular exercise,blood glucose monitoring,medication compliance and SDSCA total score(P<0.05),and indirect and total effects on regular diet score(P<0.05).The mediating effect of empowerment ability was significant(Bootstrap CI did not include 0).The chain mediating effect analysis showed that type D personality could indirectly affect HbA1c through empowerment ability,healthy diet(γ=0.389,95%CI 0.206～0.591),and medication compliance(γ=0.149,95%CI 0.040～0.265),and the effect proportion was 39.4%and 14.1%,respectively.Conclusions Type D personality can indirectly influence self-management behavior through the mediating effect of empowerment,and simultaneously affecting HbA1c through the chain effect of empowerment,diet,and medication behavior.

Objective:To investigate the characteristics of the endometrial microbiota in patients with varying degrees of intrauterine adhesion (IUA).Methods:This single-center cross-sectional observational study enrolled 115 patients with IUA who were treated at the Hysteroscopic Center of Fuxing Hospital, Capital Medical University, from May 2022 to October 2023. After quality control and data preprocessing, 81 samples met the inclusion criteria for analysis. Patients were grouped according to an established IUA scoring and grading system into mild IUA ( n=38) and moderate-to-severe IUA ( n=43). Endometrial tissue was collected under sterile conditions. Bacterial genomic DNA was extracted, the 16S rRNA V3-V4 region was amplified, and sequencing was performed on an Illumina platform. Differences in endometrial microbiota diversity and composition were compared between the two groups. Results:Patients with varying degrees of IUA exhibited comparable species richness, evenness and diversity of endometrial microbiota. At the phylum level, the endometrial microbiota across all subjects was predominantly composed of Proteobacteria, Firmicutes, Cyanobacteriota, Bacteroidota, and Actinobacteriota, with Proteobacteria (32.29%) and Firmicutes (23.82%) showing the highest mean relative abundances. At the genus level, Ralstonia (16.67%), Lactobacillus (13.45%), and Streptococcus (7.07%) were the most abundant genera. Group comparisons showed that the abundance of Ralstonia was higher in the mild IUA group, whereas Lactobacillus, Vibrio and Pseudoalteromonas were more abundant in the moderate-to-severe IUA group; however, these differences did not reach statistical significance (all P>0.05). LEfSe analysis further indicated that Lactobacillus, Vibrio, Pseudoalteromonas, Aeromonas, Ureaplasma and Acetobacterium were relatively enriched in the moderate-to-severe IUA group, while Geobacillus, Stomatobaculum and Fusicatenibacter were more abundant in the mild IUA group. Conclusion:The composition of the endometrial microbiota differs among patients with varying IUA severity. IUA progression may be associated with alterations in the endometrial microbiota; however, causal relationships and underlying mechanisms require further investigation.