ObjectiveTo observe the clinical effectiveness of horticultural therapy involving the planting of Chinese medicinal herbs （mint and lily potted plants） combined with intradermal needling therapy for generalized anxiety disorder （GAD） of liver depression transforming into fire syndrome under transcranial magnetic stimulation and basic psychological therapy， and to explore the possible mechanisms of action. MethodsA total of 180 patients with GAD of liver depression transforming into fire syndrome were randomly divided into three groups， horticultural therapy group， intradermal needling group， and horticultural therapy+intradermal needling group， with 60 patients in each. All groups received basic treatment including basic psychological therapy and transcranial magnetic stimulation. The horticultural therapy group received horticultural therapy in addition to the basic treatment； the intradermal needling group received intradermal needling therapy once a week for 8 weeks in addition to the basic treatment； the horticultural therapy+intradermal needling group received both horticultural therapy and intradermal needling therapy， following the same procedures and duration. Hamilton Anxiety Rating Scale （HAMA）， Self-Rating Anxiety Scale （SAS）， and Pittsburgh Sleep Quality Index （PSQI） scores were assessed at baseline and after 2， 4， 6， and 8 weeks of treatment. Serum levels of adrenocorticotropic hormone （ACTH） and corticosterone （CORT） were measured before treatment and after 8 weeks of treatment. Motor-evoked potential （MEP） baseline levels were recorded before treatment， and MEP amplitude ratios were compared after 1 week and 8 weeks of treatment. Clinical effectiveness and safety were evaluated after 8 weeks of treatment. Pearson correlation analysis was used to examine the relationships between serum ACTH and CORT levels， MEP amplitude， and anxiety. ResultsIn the horticultural therapy group and intradermal needling group， HAMA， SAS and PSQI scores after 4， 6， and 8 weeks treatment were lower than baseline scores （P＜0.05）. In the horticultural therapy+intradermal needling group， these scores showed a significant decline starting after 2 weeks treatment and continuing through 8 weeks after treatment （P＜0.05）. The HAMA， SAS， and PSQI scores in the horticultural therapy+intradermal needling group were significantly lower than those in the other two groups after 2， 4， 6， and 8 weeks treatment （P＜0.05）. After 8 weeks of treatment， serum CORT and ACTH levels in the horticultural therapy+intradermal needling group were significantly lower than baseline levels （P＜0.05） and were also lower than those in the horticultural therapy group and intradermal needling group at the same time point （P＜0.01）. When comparing the level after 8 weeks treatment to that after 1 week treatment， under PAS10 stimulation， the MEP amplitude ratio in the intradermal needling group decreased at 30 minutes， while in the horticultural therapy+intradermal needling group， the MEP amplitude ratio decreased at all time points （P＜0.05 or P＜0.001）； under PAS25 stimulation， the MEP amplitude ratio in the horticultural therapy group increased at 20 minutes， and in the intradermal needle group at 10 minutes （P＜0.05）. In the horticultural therapy+intradermal needling group， the MEP amplitude ratio increased significantly at all time points after treatment （P＜0.001）. The cure rate in the horticultural therapy+intradermal needling group （74.14%， 43/58） was significantly higher than that in the horticultural therapy group （30.00%， 18/60） and the intradermal needling group （48.28%， 28/58， P＜0.05）. Correlation analysis revealed that serum ACTH and CORT levels were positively correlated with HAMA scores （r = 0.488， P＜0.01； r = 0.428， P＜0.01）. Following PAS10 intervention， the MEP amplitude ratio was positively correlated with HAMA scores （r = 0.458， P＜0.01）， whereas after PAS25 intervention， the MEP amplitude ratio was negatively correlated with HAMA scores （r = -0.562， P＜0.01）. ConclusionHorticultural therapy combined with intradermal needling treatment， under transcranial magnetic stimulation and basic psychological therapy， demonstrates significant clinical effectiveness in patients with GAD of liver depression transforming into fire syndrome. Its mechanism of action may be related to the regulation of hyperactivation of the hypothalamic-pituitary-adrenal （HPA） axis and the reduction of cortical excitability.

OBJECTIVES: Cardiovascular disease (CVD) poses a major threat to global health. Evaluating atherosclerosis in asymptomatic individuals can help identify those at high risk of CVD. This study aims to establish an individualized nomogram prediction model to estimate the risk of vascular plaque formation in asymptomatic individuals. METHODS: A total of 5 655 participants who underwent CVD screening at the Health Management Center of The Third Xiangya Hospital, Central South University, between January 2022 and June 2024 we retrospectively enrolled. Using simple random sampling, participants were divided into a training set (n=4 524) and a validation set (n=1 131) in an 8꞉2 ratio. Demographic and clinical data were collected and compared between groups. Multivariate logistic regression analysis was used to identify independent factors associated with vascular plaques and to construct a nomogram prediction model. The predictive performance and clinical utility of the model were evaluated using receiver operating characteristic (ROC) curves, the Hosmer-Lemeshow goodness-of-fit test, calibration plots, and decision curve analysis (DCA). RESULTS: The mean age of participants was 52 years old. There were 3 400 males (60.12%). The overall detection rate of vascular plaque in the screening population was 49.87% (2 820/5 655). No statistically significant differences were observed in clinical indicators between the training and validation sets (all P>0.05). Multivariate Logistic regression analysis identified age, systolic blood pressure, high-density lipoprotein (HDL), low-density lipoprotein (LDL), lipoprotein(a), male sex, smoking history, hypertension history, and diabetes history as independent risk factors for vascular plaque in asymptomatic individuals (all P<0.05). The area under the curve (AUC) of the nomogram model for predicting vascular plaque risk were 0.778 (95% CI 0.765 to 0.791, P<0.001) in the training set and 0.760 (95% CI 0.732 to 0.787, P<0.001) in the validation set. The Hosmer-Lemeshow goodness-of-fit test indicated good model calibration (training set: P=0.628; validation set: P=0.561). The calibration curve plotted using the Bootstrap method demonstrated good agreement between predicted probabilities and actual probabilities. DCA showed that the nomogram provided a clinical net benefit for predicting vascular plaque risk when the threshold probability ranged from 0.02 to 0.99. CONCLUSIONS The nomogram prediction model for vascular plaque risk, constructed using readily available and cost-effective physical examination indicators, exhibited good predictive performance. This model can assist in the early identification and intervention of asymptomatic individuals at high risk for cardiovascular disease.
Humans ; Male ; Middle Aged ; Female ; Nomograms ; Retrospective Studies ; Risk Factors ; Plaque, Atherosclerotic/diagnosis* ; Aged ; Adult ; Physical Examination ; Logistic Models ; Cardiovascular Diseases/epidemiology* ; ROC Curve

OBJECTIVE: This study aims to elucidate the therapeutic potential of osthole for the treatment of atopic dermatitis (AD), focusing on its ability to alleviate chronic pruritus (CP) and the underlying molecular mechanisms. METHODS: In this study, we investigated the anti-inflammatory effects of osthole in both a 2,4-dichloronitrobenzene (DNCB)-induced AD mouse model and tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) stimulated huma immortalized epidermal (HaCaT) cells. The anti-itch effect of osthole was specifically assessed in the AD mouse model. Using methods such as hematoxylin and eosin (HE) staining, enzyme-linked immunosorbent assay (ELISA), western blot (WB), quantitative real-time PCR (qRT-PCR), and immunofluorescence staining. RESULTS: Osthole improved skin damage and clinical dermatitis scores, reduced scratching bouts, and decreased epidermal thickness AD-like mice. It also reduced the levels of interleukin (IL)-31 and IL-31 receptor A (IL-31 RA) in both skin tissues and HaCaT cells. Furthermore, Osthole suppressed the protein expression levels of phosphor-p65 (p-p65) and phosphor-inhibitor of nuclear factor kappa-Bα (p-IκBα). Meanwhile, it increased the protein expression levels of peroxisome proliferator-activated receptor α (PPARα) and PPARγ in HaCaT cells. CONCLUSION These findings indicated that osthole effectively inhibited CP in AD by activating PPARα, PPARγ, repressing the NF-κB signaling pathway, as well as the expression of IL-31 and IL-31 RA.

Transaminases are a class of enzymes that catalyze the transfer of amino between amino acids and keto acids, playing an important role in the biosynthesis of organic amines and the corresponding derivatives. However, natural enzymes often have low catalytic efficiency against non-natural substrates, which limits their widespread applications. Enzyme engineering serves as an effective approach to improve the catalytic properties and thereby expand the application scope of transaminases. In this study, a high-throughput screening method for transaminases was established based on the fluorescent color reaction between methoxy-2-aminobenzoxime (PMA) and ketones. According to the changes in fluorescence intensity, the concentration changes of ketones could be easily monitored. The efficiency, sensitivity, and accuracy of the screening method were improved by optimization of the system. With 4-hydroxy-2-butanone as the substrate, the mutant library of the transaminase from Actinobacteria sp. was established and a mutant with increased activity was successfully obtained, which improved the production efficiency of (R)-3-aminobutanol by enzyme-catalyzed synthesis. This study laid an important foundation for efficient screening, modification, and application of transaminase.
Transaminases/metabolism* ; Fluorescent Dyes/chemistry* ; High-Throughput Screening Assays/methods* ; Ketones/metabolism* ; Actinobacteria/enzymology*

Objective To investigate the role of exosomal microRNA(miRNA)miR-210-3p in chemoresistance and stem cell property formation in cervical cancer,and to elucidate its underlying mechanism through targeting of F-box protein 31(FBXO31).Methods Exosomes were isolated from cisplatin-sensitive HeLa cells and cisplatin-resistant HeLa/DDP cells via ultracentrifugation,and their uptake by HeLa/DDP cells was verified using the PKH26 red fluorescent labeling method.HeLa cells were transfected with NC inhibitor,miR-210-3p inhibitor alone,or in combination with si-NC and si-FBXO31.After 24 hours of transfection,exosomes were extracted and co-cultured with HeLa/DDP cells for 48 hours.Consequently,HeLa/DDP cells were divided into five groups:the Ctrl group(PBS blank control),the NC inhibitor group,the miR-210-3p inhibitor group,the miR-210-3p inhibitor+si-NC group and the miR-210-3p inhibitor+si-FBXO31 group.RT-qPCR was used to measure miR-210-3p and FBXO31 expression levels.The half-maximal inhibitory concentration(IC50)of cisplatin was determined using the MTT assay.Stem cell properties were assessed via tumor sphere formation assays.Western blot analysis was performed to detect the protein expression of FBXO31 and stem cell markers(e.g.,SOX2,OCT4,NANOG).The targeting relationship between miR-210-3p and FBXO31 was validated using dual-luciferase reporter assays.The effect of exosomal miR-210-3p on the metastasis of cervical cancer in vivo was evaluated by nude mice xenograft tumor.Resuts Compared with human normal cervical epithelial cells(HCeEpiC),miR-210-3p expression was significantly upregulated in cervical cancer cell lines(HeLa,HT3,C33A,and CaSki),while FBXO31 expression was significantly downregulated(P<0.05).HeLa/DDP cells(cisplatin-resistant)exhibited significantly higher miR-210-3p expression levels and IC50 values for cisplatin compared with parental HeLa cells(P<0.05),and HeLa exosomes were efficiently taken up by HeLa/DDP cells.Compared with the NC inhibitor groupThe miR-210-3p inhibitor group showed significantly reduced expression levels of miR-210-3p,OCT4,SOX2,and NANOG,as well as a significantly lower IC50(P<0.05),while FBXO31 expression was significantly increased(P<0.05).Compared with the miR-210-3p inhibitor+si-NC group,the miR-210-3p inhibitor+si-FBXO31 group exhibited significantly decreased FBXO31 expression(P<0.05)and increased IC50,Oct-4,SOX2,and NANOG expression(P<0.05).Compared with the control group and the empty vector group,the tumor weight and volume were significantly lower in the miR-210-3p than in the control and empty vector groups(P<0.05).Conclusion Exosomal miR-210-3p promotes chemoresistance to DDP and enhances stem cell-like properties in CC cells by directly targeting and inhibiting FBXO31.

BACKGROUND:Frog active peptides have rich activities,such as antibacterial and anti-tumor,and are expected to solve the problem of antibiotic resistance. OBJECTIVE:The active peptide QUB2984 was discovered in the skin secretions of Agalychnis callidryas.Its structure and properties were simulated by bioinformatics.The peptide was synthesized,purified,and identified and its biological functions were investigated. METHODS:Agalychnis callidryas skin secretions were collected by electrostimulation.The sequence of QUB2984 was obtained through constructing a cDNA library with isolated mRNA.BLAST was used for peptide sequence alignment.Besides that,Iterative Threading ASSEmbly Refinement(I-TASSER)and HeliQuest tools were used for protein secondary structure simulation.It was synthesized by solid-phase peptide synthesis,purified by reverse-phase high-performance liquid chromatography,and structurally confirmed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.The purified peptide was used to evaluate its biological activity.Its antibacterial effect was evaluated by the minimum inhibitory concentration method.Its cytotoxic effect was detected by MTT assay.Its safety was investigated by a hemolysis test. RESULTS AND CONCLUSION:(1)Peptide QUB2984 had basically α-spiral structure,with a relatively intact hydrophobic surface,and a certain destructive ability to biofilm.The third amino acid position of QUB2984 was composed of W and had a G-X-G structure.(2)The minimum inhibitory concentration of QUB2984 against gram-positive Staphylococcus aureus was 2 μmol/L,the minimum inhibitory concentration against gram-negative Escherichia coli was 2 μmol/L,and the minimum inhibitory concentration against the fungus Candida albicans was 8 μmol/L.(3)The active peptide QUB2984 had obvious inhibitory effect on human non-small cell lung cancer cells NCI-H838 at 10-5 mol/L concentration,and the hemolytic effect on horse red cells at 64 μmol/L concentration was 50%.(4)The results showed that QUB2984 had anti-bacterial and anti-cancer activity,and it had a positive charge of +3,which was conducive to contact with bacteria or cells.

Objective Given that the PI3K/AKT/mTOR signaling pathway is associated with the progression of knee osteoarthritis(KOA),this study aims to investigate whether the polarization induction of synovial macrophages mediated by the PI3K/AKT/mTOR signaling axis is the cause of KOA progression.Methods The synovial fluid of KOA KL-Ⅱ and KL-Ⅲ patients and normal individuals was collected,and the percentage of M1 macrophages(CD80,CD86)and M2 macrophages(CD163,CD206)in the synovial fluid(M1/M2 ratio)was measured to e-valuate the polarization of macrophage cytokines such as IL-1,IL-6,IL-10,and tumor necrosis factor(TNF)-α,transforming growth factor(TGF)-β Expression in KOA synovial fluid,and detect and analyze of key molecules PI3K/AKT/mTOR signaling axis PI3K,AKT3,mTORC1,and inducible nitric oxide synthase(iONS)in KOA synovial fluid.Results Compared with the synovial fluid of normal individuals,the percentage of M1 macrophages(CD80,CD86)in KOA patients increased(P<0.01),and the M1/M2 ratio increased(P<0.001);The ex-pression of IL-1,IL-6,and TNF-α in the synovial fluid of the KOA group was also higher than that of the control group(P<0.01),while the expression of IL-10 and TGF-β in the KOA group was significantly reduced(P<0.01);The key proteins PI3K,AKT3,mTORC1,and downstream inflammatory factor iONS in the PI3K/AKT/mTOR signaling pathway in the synovial fluid of the KOA group were higher than those in the control group(P<0.01).Conclusion In KOA synovial fluid,M1 macrophage polarization plays a dominant role,and the inflam-matory response mediated by M1 macrophage polarization may be the cause of synovitis.At the same time,the PI3K/AKT/mTOR signaling pathway may mediate the polarization of M1 macrophages involved in KOA inflammato-ry response.

Objective:To investigate the relationship between different obesity metabolic phenotypes and the incidence of new carotid artery plaque.Methods:The present study is a retrospective cohort study, collecting individuals from the Health Management Center of the Second Affiliated Hospital of Dalian Medical University who had two or more cervical vascular color ultrasound examinations and met the inclusion criteria from 2014 to 2022, and collected their baseline clinical data. According to whether the subjects were obese and had metabolic syndrome, they were divided into metabolically healthy non-obese group, metabolically unhealthy non-obese group, metabolically healthy obese group, and metabolically unhealthy obese group. The first physical examination time of the subjects was taken as the starting point of follow-up, and cervical vascular color ultrasound was performed during the follow-up physical examination, with the outcome event being carotid artery plaque. Kaplan-Meier survival curve analysis was used to analyze the cumulative incidence of carotid artery plaques in the four groups and log-rank test was performed, and a multifactorial Cox proportional hazards model was used to analyze the relationship between different obesity metabolic phenotypes and the risk of carotid artery plaque incidence.Results:A total of 4 890 subjects were enrolled, aged (45.4±9.6) years, and 2 754 (56.3%) males. The follow-up time was 1.14(0.93, 2.20) years. Compared with the other 3 obesity metabolic phenotypes, the incidence of carotid plaques in the metabolically unhealthy obesity group was the highest (15.4% (286/1 861)). Kaplan-Meier survival curve analysis showed that the cumulative incidence of carotid plaques in metabolically unhealthy obese subjects was about 2.962 times that of metabolically healthy non-obese subjects (log-rank P<0.001). Multivariate Cox regression results showed that the risk of carotid plaque in metabolically unhealthy obese subjects was 1.650 times that of metabolically healthy non-obese subjects (95% CI: 1.203-2.264, P=0.002). Conclusion:Metabolically unhealthy obesity phenotype is an independent risk factor for carotid plaque.

Objective To investigate the relationship between enlarged perivascular spaces in the hippocampus(H-EPVS)and cognitive function in patients with type 2 diabetes mellitus(T2DM),and to explore whether the H-EPVS can be used as an imaging marker of cognitive impairment in T2DM.Methods A total of 93 T2DM patients were selected(including 56 patients with cognitive impair-ment and 37 patients with normal cognition).All patients received head MRI scanning and neuropsychological tests.The differences of the H-EPVS counts and various parameters between the two groups were compared.Multiple linear regression was used to evalu-ate the association between H-EPVS and cognitive function.Results Compared with the T2DM patients with normal cognition,the cognitive impairment had significantly higher total H-EPVS counts(P=0.015),and worse performance of global cognitive function,processing speed,immediate memory and delayed recall(P＜0.05).After adjusting for confounding factors by multiple linear regres-sion,total H-EPVS were associated with global cognitive function and immediate memory.Conclusion The H-EPVS are correlated with global cognitive function and memory function in patients with T2DM.The H-EPVS may be used as an imaging marker of cog-nitive impairment in T2DM patients.

Objective:To study the correlation between different body weight metabolic phenotypes and their changes and new-onset hyperuricemia in physical examination population.Methods:This study was a retrospective cohort study. A total of 31 956 people who underwent routine physical examination and met the inclusion and exclusion criteria at the Health Management Center of the Second Affiliated Hospital of Dalian Medical University from January 1, 2014 to August 31, 2022 were selected as the study subjects to establish a dynamic physical examination cohort. The end point of follow-up was new-onset hyperuricemia or the end of follow-up period. Cox regression stepwise fitting model was used to analyze the risk of different body weight metabolic phenotypes and hyperuricemia, and stratified analysis was performed for gender. According to body weight metabolic phenotype, the subjects were divided into normal metabolism and normal weight(NMNW) group, normal metabolism and obesity (NMO) group, abnormal metabolism and normal weight (AMNW) group and abnormal metabolism and obesity (AMO) group. The risk of hyperuricemia was calculated according to the changes of body weight metabolic phenotype during the follow-up period. In the sensitivity analysis, the robustness of the results was verified by changing the diagnostic criteria for hyperuricemia, removing patients with hyperuricemia at the first year of follow-up, and removing subjects aged ≥65 years.Results:Compared with the NMNW group, the risk of hyperuricemia in the NMO group, AMNW group and AMO group increased by 78.9%, 61.3%, 115.4%, respectively ( χ2=272.88, 128.15, 496.12, all P<0.001). Patients who were initially classified as NMNW at baseline, if transitioned to NMO or AMO by the follow-up endpoint, their risk of hyperuricemia increased by 122.5% ( χ2=8.01, P<0.05) and 137.4% ( χ2=15.99, P<0.001), respectively. When the baseline AMNW group changed to AMO, the risk of hyperuricemia was increased by 119.2% ( χ2=6.63, P<0.05). For patients with AMO as baseline, if they turned into NMNW and AMNW at the end of follow-up, their risk of hyperuricemia would decrease by 58.3% ( χ2=43.67, P<0.001) and 27.2% ( χ2=16.07, P<0.001). Patients with a baseline of NMO who transitioned to NMNW and AMNW at the follow-up endpoint had their risk of developing hyperuricemia decreased by 36.7% ( χ2=25.35, P<0.001) and 30.9% ( χ2=9.70, P<0.05), respectively. Conclusions:The transition from metabolic health and non-overweight obesity to metabolic abnormalities and overweight obesity is associated with an increased risk of hyperuricemia, and improvements in metabolic health or weight are associated with a decreased risk of hyperuricemia.