BACKGROUND:Naringin has been shown to promote the proliferation and osteogenic differentiation of bone marrow derived mesenchymal stem cells,making it a potential candidate for treating osteoporosis andenhancing fracture healing.However,its clinical application is limited by its low bioavailability.OBJECTIVE:To prepare chitosan/β-tricalcium phosphate scaffolds loaded with naringin and characterize their biological properties.METHODS:Chitosan/β-tricalcium phosphate scaffolds were prepared by freeze-drying and chemical crosslinking.The chitosan/β-tricalcium phosphate scaffolds were immersed in anhydrous ethanol solution containing naringin for 3 hours.After vacuum cold drying,chitosan/β-tricalcium phosphate/naringin scaffolds were obtained.The pore size,porosity,swelling rate,degradation rate,mechanical properties,and in vitro release capacity of naringin of the scaffolds were characterized.Rat bone marrow mesenchymal stem cells were inoculated on the surface of chitosan/β-tricalcium phosphate scaffolds and chitosan/β-tricalcium phosphate/naringin scaffolds,respectively,and cell proliferation,adhesion,activity and alkaline phosphatase activity after osteogenic differentiation were detected.RESULTS AND CONCLUSION:(1)The results of scanning electron microscopy showed that the naringin-chitosan/β-tricalcium phosphate composite scaffold had a porous mesh structure.The average pore diameter was(106.82±25.22)μm;the porosity was(76.26±4.81)％;24-hour swelling rate was(796.17±31.76)％;in vitro degradation rate of 7.71％at 4 weeks,and naringin could be slowly released in vitro for 9 days.There was no significant difference in the average pore size,porosity,24-hour swelling rate,in vitro degradation rate,compression strength and compression modulus at 4 weeks between the chitosan/β-tricalcium phosphate scaffold and the chitosan/β-tricalcium phosphate scaffold(P＞0.05).(2)Rat bone marrow mesenchymal stem cells adhered well to the surfaces of the two scaffolds and had good activity.Compared with the chitosan/β-tricalcium phosphate scaffold,the chitosan/β-tricalcium phosphate/naringin scaffold promoted the proliferation of rat bone marrow mesenchymal stem cells(P＜0.05),and increased the alkaline phosphatase activity of bone marrow mesenchymal stem cells after osteogenic differentiation(P＜0.05).(3)The results show that the chitosan/β-tricalcium phosphate/naringin scaffolds exhibit favorable physical properties and can effectively promote the adhesion,proliferation,and osteogenic differentiation of bone marrow-derived mesenchymal stem cells.

BACKGROUND:Naringin has been shown to promote the proliferation and osteogenic differentiation of bone marrow derived mesenchymal stem cells,making it a potential candidate for treating osteoporosis andenhancing fracture healing.However,its clinical application is limited by its low bioavailability.OBJECTIVE:To prepare chitosan/β-tricalcium phosphate scaffolds loaded with naringin and characterize their biological properties.METHODS:Chitosan/β-tricalcium phosphate scaffolds were prepared by freeze-drying and chemical crosslinking.The chitosan/β-tricalcium phosphate scaffolds were immersed in anhydrous ethanol solution containing naringin for 3 hours.After vacuum cold drying,chitosan/β-tricalcium phosphate/naringin scaffolds were obtained.The pore size,porosity,swelling rate,degradation rate,mechanical properties,and in vitro release capacity of naringin of the scaffolds were characterized.Rat bone marrow mesenchymal stem cells were inoculated on the surface of chitosan/β-tricalcium phosphate scaffolds and chitosan/β-tricalcium phosphate/naringin scaffolds,respectively,and cell proliferation,adhesion,activity and alkaline phosphatase activity after osteogenic differentiation were detected.RESULTS AND CONCLUSION:(1)The results of scanning electron microscopy showed that the naringin-chitosan/β-tricalcium phosphate composite scaffold had a porous mesh structure.The average pore diameter was(106.82±25.22)μm;the porosity was(76.26±4.81)％;24-hour swelling rate was(796.17±31.76)％;in vitro degradation rate of 7.71％at 4 weeks,and naringin could be slowly released in vitro for 9 days.There was no significant difference in the average pore size,porosity,24-hour swelling rate,in vitro degradation rate,compression strength and compression modulus at 4 weeks between the chitosan/β-tricalcium phosphate scaffold and the chitosan/β-tricalcium phosphate scaffold(P＞0.05).(2)Rat bone marrow mesenchymal stem cells adhered well to the surfaces of the two scaffolds and had good activity.Compared with the chitosan/β-tricalcium phosphate scaffold,the chitosan/β-tricalcium phosphate/naringin scaffold promoted the proliferation of rat bone marrow mesenchymal stem cells(P＜0.05),and increased the alkaline phosphatase activity of bone marrow mesenchymal stem cells after osteogenic differentiation(P＜0.05).(3)The results show that the chitosan/β-tricalcium phosphate/naringin scaffolds exhibit favorable physical properties and can effectively promote the adhesion,proliferation,and osteogenic differentiation of bone marrow-derived mesenchymal stem cells.

ObjectiveTo explore the effects of hypoxia on the migration ability of human oligodendrocyte precursor cells （hOPCs） and its regulatory mechanisms. MethodsBased on the variations in oxygen concentration within the culture system， three experimental groups were set up： the 21%O₂ group （normoxic control group）， the 5%O₂ group， and the 2%O₂ group. The migration ability of hOPCs under normoxia （21%O₂）， 5%O₂， and 2%O₂ conditions was detected through the Transwell migration assay. RT-qPCR， transcriptome sequencing， and flow cytometry were used to detect the expression changes of genes and proteins such as hypoxia-inducible factor 1 alpha （HIF-1α） and chemokine （C-X-C Motif） receptor 4 （CXCR4）. Bioinformatics analysis was combined to analyze the KEGG pathways related to migration， so as to explore the effects of different oxygen concentrations on the migration ability of hOPCs and their possible mechanisms. ResultsHypoxia treatments at concentrations of 5%O₂ and 2%O₂ could both promote the in vitro migration of hOPCs， and the promoting effect of migration was more significant at the 2%O₂ concentration （P<0.001）. After hypoxia treatment， the mRNA expression levels of HIF-1α， CXCR4， etc. in hOPCs significantly increased （P<0.001）. Compared with the 5%O₂ concentration， the expression of CXCR4 in cells was higher at the 2%O₂ concentration （P<0.000 1）. Flow cytometry analysis detection showed that the expression of CXCR4 increased significantly after hypoxia treatment （P<0.01）， and with the decrease of oxygen concentration， its expression level further increased （P<0.000 1）. Ordinary transcriptome sequencing analysis indicated that hypoxia treatment could activate the PI3K-Akt signaling pathway and the Axon guidance pathway. ConclusionHypoxia treatment can enhance the in vitro migration ability of hOPCs， and this effect is negatively correlated with the oxygen concentration. Its mechanism may be related to the up-regulation of the expression of genes such as HIF-1α and CXCR4， and the activation of the migration related signaling pathway including PI3K-Akt signaling pathway and axon guidance pathway.

OBJECTIVE To investigate the efficacy and safety of Wuling capsules combined with fluoxetine in the treatment of adolescents with first-episode moderate-to-severe depressive disorder accompanied by insomnia. METHODS The clinical data of 476 adolescents with first-episode moderate-to-severe depression accompanied by insomnia admitted to our hospital from June 2022 to May 2025， were retrospectively collected. According to the initial treatment regimen， patients were divided into a control group （241 cases， treated with fluoxetine alone） and an observation group （235 cases， treated with Wuling capsules combined with fluoxetine）. The depression severity （Hamilton Depression Rating Scale-17 Item and the Self-Rating Depression Scale scores）， sleep quality （Pittsburgh Sleep Quality Index score， sleep latency， wake after sleep onset， total sleep time， sleep efficiency）， serum neuroendocrine indicator （cortisol） and inflammatory markers （C-reactive protein， interleukin-6） were compared between the two groups before treatment and at 4th and 8th weeks of treatment. The effective rate at 8th weeks and the occurrence of adverse drug reactions （ADRs） were also compared between the two groups. RESULTS Before treatment， there were no significant differences in depression severity， sleep quality， serum neuroendocrine indicator， and inflammatory markers between the two groups （ P ＞0.05）. At 4th and 8th weeks， both groups showed significant improvement in these indicators compared to those before treatment， with the observation group demonstrating significantly greater improvement than the control group at the corresponding time points （ P ＜0.05）. At 8th week， the eff ective rate of the observation group was 90.21%， significantly higher than 80.50% in the control group （ P ＜0.05）. The incidence of nausea， headache， fatigue， dry mouth， and palpitations， as well as the total incidence of ADRs， did not differ significantly between the two groups （ P ＞0.05）. CONCLUSIONS Wuling capsules combined with fluoxetine can significantly improve the effective rate in adolescents with first-episode moderate-to-severe depression accompanied by insomnia， accelerate the relief of depressive symptoms， improve sleep quality， and reduce serum neuroendocrine indicator and inflammatory markers， with a favorable safety profile.

ObjectiveTo develop an assessment tool for apraxia of speech (AOS) of Chinese Putonghua speakers and test its reliability and validity. MethodsThe Chinese Apraxia of Speech Assessment and Diagnostic Tool (CAADT) was developed based on the Apraxia of Speech Rating Scale 3.5, combined with the linguistic characteristics of Chinese and clinical experience. The tool consistsed of eleven items across three sections: articulation, prosody and alternating motion rates. Six experts evaluated the content validity. From November, 2024 to May, 2025, 51 patients with post-stroke AOS (experimental group) and ten patients with post-stroke aphasia without AOS (control group) were recruited from Anhui Wannan Rehabilitation Hospital (the Fifth People's Hospital of Wuhu), and tested with CAADT. Reliability was assessed using Cronbach's α coefficient, Kendall's coefficient of concordance W and Pearson correlation coefficient. Validity was evaluated using the content validity index (CVI) and Spearman correlation coefficient. Discriminative effect was analyzed using the receiver operating characteristic (ROC) curve. ResultsThe Cronbach's α coefficients for the articulation and prosody sections and the total scale were all > 0.9, while it was 0.454 for the alternating motion rates. Inter-rater reliability was good (W ≥ 0.598, P < 0.001). Test-retest reliability showed high positive correlations for the three sections and the total score between the two assessments (r ≥ 0.84, P < 0.001). The scale-level CVI was 0.95, and the item-level CVI ≥ 0.83. The Spearman correlation coefficients among the sections ranged from 0.30 to 0.70. ROC analysis revealed an area under the curve of 0.953, with a cut-off value of 11, yielding a sensitivity of 0.92 and a specificity of 0.90. ConclusionCAADT demonstrates good reliability, validity and discriminative effect, which can be used for clinical assessment and auxiliary diagnosis of Chinese Putonghua speaking patients with post-stroke AOS.

AIM: To explore the clinical therapeutic effect of teprotumumab combined with glucocorticoid pulse therapy for thyroid-associated ophthalmopathy(TAO), and its impacts on thyroid function, levels of inflammatory factors, and adverse reactions in patients. METHODS: Active TAO patients admitted to the Ophthalmology Department were enrolled and randomly divide into the steroid group and the combined group. Then the steroid group was treated with glucocorticoid pulse therapy, while the combined group was combined with intravenous infusion of teprotumumab on the basis of the steroid group. The clinical therapeutic effect, the CAS, OSDI, M-C-TAO-QOL scores, ocular sign indicators(fissure width, proptosis), levels of inflammatory factors(TNF-α, CRP, IL-17), thyroid function(TSH, FT3, FT4)before and after treatment, and occurrence of adverse reactions were compared between two groups.RESULTS:Totally 96 TAO patients(192 eyes)were included, with 48 cases(96 eyes)in each group. In the combined group, there were 17 males and 31 females, with an average age of 51.85±3.53 y; in the steroid group, there were 14 males and 34 females, with an average age of 51.26±3.84 y. The total effective rate of the combined group(94%)was higher than that of the steroid group(79%)(P<0.05). After treatment, the CAS score, OSDI score, fissure width, proptosis, levels of TNF-α, CRP, and IL-17 in the combined group were all lower than those in the steroid group, and the M-C-TAO-QOL score was higher than that in the steroid group(P<0.05). However, there was no difference in thyroid function indicators and adverse reactions between two groups after treatment(P>0.05). CONCLUSION: The combination of teprotumumab and glucocorticoid pulse therapy for TAO has a prominent therapeutic effect. Meantime, it can more effectively control ocular inflammation, improve ocular signs and quality of life of patients, and has no obvious adverse effect on thyroid function, with controllable safety.

Objective: To address the challenges of systematically identifying herb pairs in traditional Chinese medicine (TCM), we proposed HerbGL, a framework for predicting potential herb pairs that integrates network propagation and graph regularization. Methods: Based on the assumption that herbal actions induce subtle perturbations in biological systems, a framework named HerbGL was proposed. Random walk with restart (RWR) was first applied to the protein-protein interaction (PPI) network to reconstruct herb-specific perturbation effects and generate weighted subnetworks. Then, to quantify affinity between herb pairs, two network-proximity metrics, Closeness and PageRank, were computed from the weighted subnetworks to construct herb-pair affinity matrices. Finally, these matrices, together with known herb pairs (derived from co-occurrence analysis of TCM formulas with a threshold determined from the inflection point of the frequency distribution), were incorporated into a graph regularization model to predict potential herb pairs. Model performance was assessed through baseline comparison, ablation and robustness experiment under different ratios of positive and negative samples, using the area under the receiver operating characteristic curve (AUROC), the area under the precision-recall curve (AUPRC), accuracy, and precision as evaluation metrics. Furthermore, the predicted herb pairs were validated through both literature evidence and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. Results: The weighted subnetworks constructed by RWR provided a refined simulation of herb-specific perturbation effects, which formed the basis for subsequent affinity modeling and prediction. Analysis of herb pair co-occurrence frequencies revealed a marked change around 150, which was selected as the threshold to distinguish herb pairs from non-herb pairs. HerbGL exhibited superior predictive performance compared with baseline models (AUROC = 0.970 5, AUPRC = 0.955 5, accuracy = 0.726 6, precision = 0.970 6). Ablation results showed that removing the Closeness and PageRank metrics substantially degraded performance (AUROC = 0.819 1, AUPRC = 0.876 8), confirming their necessity. Robustness evaluation under an imbalanced positive-to-negative sample ratio of 1 : 5 yielded AUROC = 0.969 6 and AUPRC = 0.840 4, indicating stable predictive ability. Moreover, multiple case studies further validated the rationality of the predicted herb pairs, such as Fangfeng (Saposhnikoviae Radix) and Qingpi (Citri Reticulatae Pericarpium Viride) which are recorded in Liangpeng Huiji (《良朋汇集》, Collection of Excellent Recipes) Vol. 3: Fangfeng Shengma Tang (防风升麻汤). Additionally, pathway enrichment analysis of the Renshen (Ginseng Radix et Rhizoma) and Lianqiao (Forsythiae Fructus) pair further supported the biological plausibility of their compatibility. Conclusion HerbGL offers an effective and biologically informed framework for identifying herb pairs in TCM. Beyond improving herb pair prediction, the framework also provides data support for research on herb compounds and mechanisms, thereby supporting data-driven exploration of TCM compatibility.

AIM: To investigate the effects of high-concentration glucose on the cell viability of retinal pigment epithelialcells(ARPE-19)/ human umbilical vein endothelial cells(HUVEC)cells, as well as on the expression levels of sirtuin 6(SIRT6).

METHODS:ARPE-19 and HUVEC cells were cultured for 48 h in media containing various glucose concentrations(5.5, 25, 50, 75, 100 mmol/L)and cellular morphology was observed under an optical microscope.Cell viability was assessed using the CCK-8 assay. Flow cytometry was utilized to evaluate apoptosis levels, and Western blot analysis was conducted to measure the expression of Bcl-2 and SIRT6. ARPE-19/HUVEC cells were treated with 50 mmol/L glucose for 48 h, after which immunofluorescence staining was performed to determine the subcellular localization and expression level of SIRT6 protein. In parallel, cells receiving the same treatment were harvested at different time points(0, 6, 12, 24, 36, 48, and 72 h), and the temporal changes in SIRT6 protein expression were assessed by Western blot analysis. SIRT6 overexpressing stable ARPE-19/HUVEC cell lines were constructed. Western blot was performed to detect the effects of SIRT6 overexpression on the expression levels of glucose transporters in ARPE-19/HUVEC cells treated with high glucose(50 mmol/L glucose).

RESULTS:Compared with the 5.5 mmol/L glucose group, cell number was decreased and cell viability was significantly reduced in ARPE19/HUVEC cells with increasing glucose concentrations(all P<0.01). Meanwhile, the protein expression level of SIRT6 was markedly downregulated(P<0.05). Such inhibitory effects exhibited both concentration-and time-dependent patterns(all P<0.05). Furthermore, high glucose induced cell apoptosis and downregulated the protein expression of Bcl-2(P<0.05). Immunofluorescence staining revealed that SIRT6 protein was predominantly localized in the nucleus after 48 h of high-glucose treatment, accompanied by pyknosis and nuclear fragmentation. After incubation with highg-lucose medium(50 mmol/L glucose)for different durations, long-term high-glucose intervention was found to reduce SIRT6 protein expression in ARPE19/HUVEC cells. Successful construction of the SIRT6-overexpressing stable ARPE-19/HUVEC cell lines was confirmed via immunofluorescence and Western blotting. Western blot analysis revealed that, compared with the HG+LV-con group, the HG+LV-SIRT6 group exhibited no significant differences in GLUT1 or GLUT4 expression in ARPE-19 cells, whereas GLUT3 expression was elevated(P<0.05). Compared with the HG+LV-con group, the HG+LV-SIRT6 group showed no significant change in GLUT1 expression, while GLUT3 and GLUT4 levels were increased(both P<0.05).

CONCLUSION:High-concentration glucose-reduce the proliferative activity of ARPE-19/HUVEC cells, induce apoptosis, and downregulate SIRT6 expression. SIRT6 may alleviate high glucose-induced retinal cell damage by promoting the expression of glucose transporters.

Objective:To establish an atomic fluorescence detection method for bismuth and bismuth telluride (Bi 2Te 3) in the air of workplaces. Methods:In March 2024, the acetate-fiber filter material was digested by ultrawave and detected by atomic fluorescence spectrometry to obtain the content of bismuth and Bi 2Te 3 in the air of the workplace. Results:The correlation coefficients of standard curve were above 0.9990, The detection limit of bismuth was 0.02 μg/L, the minimum detection concentration was 0.02 μg/m 3, the minimum quantitation concentration was 0.06 μg/m 3, the minimum detection concentration and the minimum quantitation concentration of Bi 2Te 3 was 0.03 μg/m 3 and 0.09 μg/m 3 respectively. The digestion efficiency ranged from 98.6% to 102 %, the RSDs of intra-batch and inter-batch were below 1.3% and 4.0% respectively. The samples were stable within 7 days, and interference elements of 1.0 mg/L (Ag, Al, As, B, Ba, Be, Cd, Co, Cr, Cs, Cu, Fe, Ga, Li, Mg, Mn, Mo, Ni, Pb, Rb, Sb, Se, Sn, Sr, Ti, Tl, V, Zn, Zr) do not affect the detection of bismuth. Conclusion:This method is simple, fast, accurate, and highly sensitive, and can meet the requirements of the determination of bismuth and bismuth telluride in the air of working place.

Objective:To evaluate the application effect of a wearable visual field meter based on extended reality (XR) glasses for patients with macular disease.Methods:A self-controlled study was conducted.A total of 41 consecutive patients (41 eyes) with macular disease were recruited at Renmin Hospital of Wuhan University from October 2022 to October 2024.All patients underwent 10-2 center visual field test using a self-developed wearable visual field meter (XRVF), and the results were compared with those obtained using a traditional Humphrey field analyzer (HFA).The comparison parameters included mean retinal sensitivity (MS), false positive rate (FPR), false negative rate (FNR), and testing duration.A subject satisfaction questionnaire was administered.This study followed the Declaration of Helsinki.The study protocol was approved by the Medical Ethics Committee of Renmin Hospital of Wuhan University (No.WDRY2024-K263), and all subjects signed the informed consent form.Results:The retinal sensitivity of patients measured by HFA was (23.24±3.71)dB, which was higher than (22.01±3.45)dB by XRVF, showing a statistically significant difference ( t=4.924, P=0.036).The FPR measured by HFA and XRVF were (2.39±2.51)% and (2.59±3.29)%, respectively, and the FNR were (3.49±6.05)% and (3.74±5.38)%, respectively, showing no statistically significant difference ( t=-3.624, P=0.948; t=-1.241, P=0.519).The median test duration for HFA and XRVF was 6.15 (5.78, 6.65) and 5.98 (5.71, 6.69)minutes, respectively, without statistically significant difference ( Z=-1.987, P=0.953).92.6% of the subjects thought the device was comfortable, simple and practical. Conclusions:The XRVF has good consistency with the HFA, can effectively and reliably evaluate the visual field function of patients with macular disease, and is easily accepted by patients.