OBJECTIVE: To investigate the antioxidant and osteogenic induction capabilities of calcium phosphate nanoflowers (hereinafter referred to as nanoflowers) in vitro at different concentrations. METHODS: Nanoflowers were prepared using gelatin, tripolyphosphate, and calcium chloride. Their morphology, microstructure, elemental composition and distribution, diameter, and molecular constitution were characterized using scanning electron microscopy, transmission electron microscopy, Fourier transform infrared spectroscopy, and energy-dispersive spectroscopy. Femurs and tibias were harvested from twelve 4-week-old Sprague Dawley rats, and bone marrow mesenchymal stem cells (BMSCs) were isolated and cultured using the whole bone marrow adherent method, followed by passaging. The third passage cells were identified as stem cells by flow cytometry and then co-cultured with nanoflowers at concentrations of 0, 0.4, 0.8, 1.2, 1.6, 2.0, 2.4, 2.8, 3.2, and 3.6 mg/mL. Cell counting kit 8 (CCK-8) assay was performed to screen for the optimal concentration that demonstrated the best cell viability, which was subsequently used as the experimental concentration for further studies. After co-culturing BMSCs with the screened concentration of nanoflowers, the biocompatibility of the nanoflowers was verified through live/dead cell staining, scratch assay, and cytoskeleton staining. The antioxidant capacity was assessed by using reactive oxygen species (ROS) fluorescence staining. The in vitro osteoinductive ability was evaluated via alkaline phosphatase (ALP) staining, alizarin red staining, and immunofluorescence staining of osteocalcin (OCN) and Runt-related transcription factor 2 (RUNX2). All the above indicators were compared with the control group of normally cultured BMSCs without the addition of nanoflowers. RESULTS: Scanning electron microscopy revealed that the prepared nanoflowers exhibited a flower-like structure; transmission electron microscopy scans discovered that the nanoflowers possessed a multi-layered structure, and high-magnification images displayed continuous atomic arrangements, with the nanoflower diameter measuring (2.00±0.25) μm; energy-dispersive spectroscopy indicated that the nanoflowers contained elements such as C, N, O, P, and Ca, which were uniformly distributed across the flower region; Fourier transform infrared spectroscopy analyzed the absorption peaks of each component, demonstrating the successful preparation of the nanoflowers. Through CCK-8 screening, the concentrations of 0.8, 1.2, and 1.6 mg/mL were selected for subsequent experiments. The live/dead cell staining showed that nanoflowers at different concentrations exhibited good cell compatibility, with the 1.2 mg/mL concentration being the best (P<0.05). The scratch assay results indicated that the cell migration ability in the 1.2 mg/mL group was superior to the other groups (P<0.05). The cytoskeleton staining revealed that the cell morphology was well-extended in all concentration groups, with no significant difference compared to the control group. The ROS fluorescence staining demonstrated that the ROS fluorescence in all concentration groups decreased compared to the control group after lipopolysaccharide induction (P<0.05), with the 1.2 mg/mL group showing the weakest fluorescence. The ALP staining showed blue-purple nodular deposits around the cells in all groups, with the 1.2 mg/mL group being significantly more prominent. The alizarin red staining displayed orange-red mineralized nodules around the cells in all groups, with the 1.2 mg/mL group having more and denser nodules. The immunofluorescence staining revealed that the expressions of RUNX2 and OCN proteins in all concentration groups increased compared to the control group, with the 1.2 mg/mL group showing the strongest protein expression (P<0.05). CONCLUSION The study successfully prepares nanoflowers, among which the 1.2 mg/mL nanoflowers exhibits excellent cell compatibility, antioxidant properties, and osteogenic induction capability, demonstrating their potential as an artificial bone substitute material.
Animals ; Osteogenesis/drug effects* ; Mesenchymal Stem Cells/drug effects* ; Calcium Phosphates/pharmacology* ; Rats, Sprague-Dawley ; Rats ; Antioxidants/chemistry* ; Cells, Cultured ; Cell Differentiation/drug effects* ; Nanostructures/chemistry* ; Tissue Engineering/methods* ; Bone Marrow Cells/cytology* ; Coculture Techniques ; Tissue Scaffolds/chemistry* ; Male ; Biocompatible Materials/chemistry* ; Cell Survival ; Core Binding Factor Alpha 1 Subunit/metabolism* ; Cell Proliferation

Objective To understand the real-life experiences of patients with chronic obstructive pulmonary disease(COPD)in disease self-control and to inform clinical nursing practice.Methods From March to May 2024,a phenomenological research method was used to conduct semi-structured interviews with 15 patients with COPD who were either outpatients or inpatients in a tertiary hospital in Suzhou City,China,and the data were analyzed using the Colaizzi 7-step analysis method.Results Totally 3 themes and 9 sub-themes were extracted,namely self-control challenges due to cognitive deficits(lack of knowledge leads to ambiguous control direction;cognitive bias hinders control strategy formulation;negative perception leads to control avoidance),weakening trend of self-control behaviors(decision-making conflicts between immediate indulgence and delayed gratification;lack of reinforcement mechanisms leads to psycho-emotional depletion;external temptations lead the self to imitate bad behavior),urgent need for multiple supports to help with self-control(the desire for continuity and stability of family support,the need for professionalism and accuracy of healthcare guidance,and the expectation for rationality and matching of resource allocation).Conclusion Healthcare professionals should pay attention to the real experience of COPD patients in the process of disease self-control,help them effectively deal with the challenges of disease self-control,strengthen self-control behaviors,and satisfy their diversified needs by strengthening the support of families,professionals,and the community to improve poor outcomes and reduce the cost of healthcare services.

Objective To investigate the predictive value of serum soluble lectin-like oxidized low density lipoprotein receptor-1(sLOX-1)and chitinase-3-like protein 1(CHI3L1)for in-stent restenosis(ISR)in pa-tients with coronary heart disease(CHD)and myocardial bridge after anterior descending stent implantation.Methods A total of 80 patients with CHD and myocardial bridge who underwent anterior descending stent implantation in a hospital from May 2018 to May 2023 were included as the disease group.They were followed up for one year after surgery and separated into ISR group(n=31)and non ISR group(n=49)based on whether ISR occurred on coronary angiography examination.Another 80 CHD patients who received treatment in a hospital were selected as the control group.The levels of serum sLOX-1 and CHI3L1 were detected by en-zyme-linked immunosorbent assay.Multivariate Logistic regression was used to analyze the influencing factors of ISR after anterior descending stent implantation in patients with CHD combined with myocardial bridge,and to analyze the predictive value of serum sLOX-1 and CHI3L1 for the occurrence of ISR after anterior de-scending stent implantation in patients with CHD combined with myocardial bridge.Results Compared with the control group,the levels of serum sLOX-1 and CHI3L1 in the disease group increased,and the difference was statistically significant(P<0.05).The levels of serum sLOX-1 and CHI3L1 in the ISR group were high-er than those in the non-ISR group,and the distance of the myocardial bridge proximal to the stent was lower than that in the non-ISR group,the differences were statistically significant(P<0.05).Serum sLOX-1,CHI3L1,and the distance of the myocardial bridge proximal to the stent were the influencing factors for the occurrence of ISR after anterior descending stent implantation in patients with CHD combined with myocardi-al bridge(P<0.05).The area under the curve of the combined prediction of serum sLOX-1 and CHI3L1 for the occurrence of ISR after anterior descending stent implantation in patients with CHD complicated with my-ocardial bridge was superior to their individual predictions(Zcombination-sLOX-1=2.502,Zcombination-CHI3L1=2.028,P=0.012,0.043).Conclusion The levels of serum sLOX-1 and CHI3L1 in patients with CHD combined with myocardial bridge are significantly increased.The combined detection of the two has certain predictive value for the occurrence of ISR after anterior descending stent implantation in patients with CHD combined with myocardial bridge.

Objective:To investigate the effects of repetitive transcranial magnetic stimulation(rTMS) on the core symptoms, brain functional connectivity and activation in children with attention deficit hyperactivity disorder (ADHD) using functional near-infrared spectroscopy (fNIRS).Methods:From September 2022 to March 2024, a total of 35 children with ADHD were selected as research subjects and they were randomly divided into observation group ( n=17) and control group ( n=18). The control group received conventional rehabilitation therapy, while the observation group received rTMS therapy in addition to the conventional therapy. Both groups were treated every other day, with each course of treatment lasting four weeks, and a total of three courses of treatment were administered consecutively. The clinical symptoms of the children with ADHD were assessed using Swanson, Nolan, and Pelham-Ⅳ(SNAP-Ⅳ) before and after treatment. fNIRS was used to detect the relative concentration changes of oxyhemoglobin (HbO 2) and deoxyhemoglobin (HbR) in the prefrontal cortex under resting-state and Go/Nogo task conditions before and after treatment. Statistical analysis was performed using SPSS 26.0 software. Paired sample t-test were used for within-group comparisons, and independent sample t-test were used for between-group comparisons. Results:(1) After treatment, the scores for inattention, hyperactivity/impulsivity and oppositional defiant behavior in the two groups were significantly lower than before treatment ( t=3.51-18.86, all P<0.05). The scores for inattention, hyperactivity/impulsivity and oppositional defiant behavior in the observation group were significantly lower than those in the control group ( t=2.21, 2.03, 2.39, all P<0.05). (2) After treatment, the functional connectivity strength between all regions of interest in both groups was significantly higher than before treatment ( t=3.53-37.90, all P<0.05). The functional connectivity strength of the left dorsolateral prefrontal cortex (0.25±0.03, 0.21±0.03), right dorsolateral prefrontal cortex (0.12±0.02, 0.09±0.02), left medial prefrontal cortex (0.13±0.02, 0.10±0.01) and right medial prefrontal cortex (0.31±0.04, 0.24±0.06) in the observation group was significantly higher than those in the control group (all P<0.05). (3) In the Go/Nogo task, after treatment, the average HbO 2 concentrations in the left dorsolateral prefrontal cortex, right dorsolateral prefrontal cortex, left medial prefrontal cortex, right medial prefrontal cortex, left temporal lobe, and right temporal lobe in both groups were all higher than before treatment (all P<0.05). After treatment, the average HbO 2 concentrations in the left dorsolateral prefrontal cortex, right dorsolateral prefrontal cortex, left medial prefrontal cortex and right medial prefrontal cortex of the observation group were significantly higher than those of the control group (all P<0.05). Conclusion:rTMS therapy can improve the core symptoms of children with ADHD, which may be related to the strength of brain functional connectivity and activation of ADHD brain function by rTMS.

Objective To analyze the symptomatic improvement effects of vitamin D in children with autism spectrum disorder(ASD)based on the microbiota-gut-brain axis.Methods Seventy-two children with ASD were randomly divided into an observation group and a control group,with 36 cases in each group.Three cases dropped out in the control group.The observation group received 1200 IU/day of vitamin D supplementation in addition to conventional rehabilitation training,while the control group received only conventional rehabilitation training.The intervention lasted for 12 weeks.Assessments were conducted before and after the intervention using the childhood autism rating scale(CARS),autism behavior checklist(ABC),and repetitive behavior scale-revised(RBS-R).Resting-state functional connectivity of the brain was measured using near-infrared functional imaging,and serum levels of 25(OH)D3,inflammatory cytokines,and gut microbiota were analyzed.The differences in these indicators before and after the intervention were compared between the two groups to evaluate clinical efficacy.Results The between-group differences in pre-and post-intervention changes showed that the observation group had significantly greater improvements than the control group in the following measures:CARS scores(-5.92±1.40 vs.-2.55±1.43),RBS-R scores(-5.99±1.01 vs.-3.10±1.47),resting-state brain functional connectivity(0.19±0.15 vs.0.10±0.18),serum 25(OH)D3 levels[(34.89±8.18)ng/mL vs.(0.68±6.73)ng/mL],serum interleukin-6(IL-6)levels[(-6.60±6.07)pg/mL vs.(-0.74±9.45)pg/mL],IL-1β levels[(-2.56±1.33)pg/mL vs.(-0.04±2.13)pg/mL],and tumor necrosis factor-α(TNF-α)levels[(-4.09±3.85)pg/mL vs.(0.21±4.05)pg/mL](P<0.05).Post-intervention,significant differences in gut microbial β-diversity were observed between the two groups(R2=0.030,P=0.040,Adonis).LEfSe analysis revealed that the observation group exhibited enrichment in Clostridia(LDA=4.747,P=0.003),Clostridiales(LDA=4.747,P=0.003),Clostridiaceae(LDA=3.476,P=0.001),Lachnospiraceae(LDA=4.709,P=0.004),Odoribacteraceae(LDA=3.458,P=0.027),Odoribacter(LDA=3.458,P=0.027),Burkholderiales(LDA=3.339,P=0.038),Firmicutes(LDA=4.764,P=0.003),and Betaproteobacteria(LDA=3.338,P=0.037).Conclusion Vitamin D supplementation can modulate gut microbial diversity in children with ASD,significantly influence the abundance of specific gut microbiota,reduce systemic inflammatory cytokines,enhance brain functional connectivity,and alleviate clinical symptoms of ASD.

Objective:To investigate the effects of repetitive transcranial magnetic stimulation(rTMS) on the core symptoms, brain functional connectivity and activation in children with attention deficit hyperactivity disorder (ADHD) using functional near-infrared spectroscopy (fNIRS).Methods:From September 2022 to March 2024, a total of 35 children with ADHD were selected as research subjects and they were randomly divided into observation group ( n=17) and control group ( n=18). The control group received conventional rehabilitation therapy, while the observation group received rTMS therapy in addition to the conventional therapy. Both groups were treated every other day, with each course of treatment lasting four weeks, and a total of three courses of treatment were administered consecutively. The clinical symptoms of the children with ADHD were assessed using Swanson, Nolan, and Pelham-Ⅳ(SNAP-Ⅳ) before and after treatment. fNIRS was used to detect the relative concentration changes of oxyhemoglobin (HbO 2) and deoxyhemoglobin (HbR) in the prefrontal cortex under resting-state and Go/Nogo task conditions before and after treatment. Statistical analysis was performed using SPSS 26.0 software. Paired sample t-test were used for within-group comparisons, and independent sample t-test were used for between-group comparisons. Results:(1) After treatment, the scores for inattention, hyperactivity/impulsivity and oppositional defiant behavior in the two groups were significantly lower than before treatment ( t=3.51-18.86, all P<0.05). The scores for inattention, hyperactivity/impulsivity and oppositional defiant behavior in the observation group were significantly lower than those in the control group ( t=2.21, 2.03, 2.39, all P<0.05). (2) After treatment, the functional connectivity strength between all regions of interest in both groups was significantly higher than before treatment ( t=3.53-37.90, all P<0.05). The functional connectivity strength of the left dorsolateral prefrontal cortex (0.25±0.03, 0.21±0.03), right dorsolateral prefrontal cortex (0.12±0.02, 0.09±0.02), left medial prefrontal cortex (0.13±0.02, 0.10±0.01) and right medial prefrontal cortex (0.31±0.04, 0.24±0.06) in the observation group was significantly higher than those in the control group (all P<0.05). (3) In the Go/Nogo task, after treatment, the average HbO 2 concentrations in the left dorsolateral prefrontal cortex, right dorsolateral prefrontal cortex, left medial prefrontal cortex, right medial prefrontal cortex, left temporal lobe, and right temporal lobe in both groups were all higher than before treatment (all P<0.05). After treatment, the average HbO 2 concentrations in the left dorsolateral prefrontal cortex, right dorsolateral prefrontal cortex, left medial prefrontal cortex and right medial prefrontal cortex of the observation group were significantly higher than those of the control group (all P<0.05). Conclusion:rTMS therapy can improve the core symptoms of children with ADHD, which may be related to the strength of brain functional connectivity and activation of ADHD brain function by rTMS.

Objective To analyze the symptomatic improvement effects of vitamin D in children with autism spectrum disorder(ASD)based on the microbiota-gut-brain axis.Methods Seventy-two children with ASD were randomly divided into an observation group and a control group,with 36 cases in each group.Three cases dropped out in the control group.The observation group received 1200 IU/day of vitamin D supplementation in addition to conventional rehabilitation training,while the control group received only conventional rehabilitation training.The intervention lasted for 12 weeks.Assessments were conducted before and after the intervention using the childhood autism rating scale(CARS),autism behavior checklist(ABC),and repetitive behavior scale-revised(RBS-R).Resting-state functional connectivity of the brain was measured using near-infrared functional imaging,and serum levels of 25(OH)D3,inflammatory cytokines,and gut microbiota were analyzed.The differences in these indicators before and after the intervention were compared between the two groups to evaluate clinical efficacy.Results The between-group differences in pre-and post-intervention changes showed that the observation group had significantly greater improvements than the control group in the following measures:CARS scores(-5.92±1.40 vs.-2.55±1.43),RBS-R scores(-5.99±1.01 vs.-3.10±1.47),resting-state brain functional connectivity(0.19±0.15 vs.0.10±0.18),serum 25(OH)D3 levels[(34.89±8.18)ng/mL vs.(0.68±6.73)ng/mL],serum interleukin-6(IL-6)levels[(-6.60±6.07)pg/mL vs.(-0.74±9.45)pg/mL],IL-1β levels[(-2.56±1.33)pg/mL vs.(-0.04±2.13)pg/mL],and tumor necrosis factor-α(TNF-α)levels[(-4.09±3.85)pg/mL vs.(0.21±4.05)pg/mL](P<0.05).Post-intervention,significant differences in gut microbial β-diversity were observed between the two groups(R2=0.030,P=0.040,Adonis).LEfSe analysis revealed that the observation group exhibited enrichment in Clostridia(LDA=4.747,P=0.003),Clostridiales(LDA=4.747,P=0.003),Clostridiaceae(LDA=3.476,P=0.001),Lachnospiraceae(LDA=4.709,P=0.004),Odoribacteraceae(LDA=3.458,P=0.027),Odoribacter(LDA=3.458,P=0.027),Burkholderiales(LDA=3.339,P=0.038),Firmicutes(LDA=4.764,P=0.003),and Betaproteobacteria(LDA=3.338,P=0.037).Conclusion Vitamin D supplementation can modulate gut microbial diversity in children with ASD,significantly influence the abundance of specific gut microbiota,reduce systemic inflammatory cytokines,enhance brain functional connectivity,and alleviate clinical symptoms of ASD.

Objective To understand the real-life experiences of patients with chronic obstructive pulmonary disease(COPD)in disease self-control and to inform clinical nursing practice.Methods From March to May 2024,a phenomenological research method was used to conduct semi-structured interviews with 15 patients with COPD who were either outpatients or inpatients in a tertiary hospital in Suzhou City,China,and the data were analyzed using the Colaizzi 7-step analysis method.Results Totally 3 themes and 9 sub-themes were extracted,namely self-control challenges due to cognitive deficits(lack of knowledge leads to ambiguous control direction;cognitive bias hinders control strategy formulation;negative perception leads to control avoidance),weakening trend of self-control behaviors(decision-making conflicts between immediate indulgence and delayed gratification;lack of reinforcement mechanisms leads to psycho-emotional depletion;external temptations lead the self to imitate bad behavior),urgent need for multiple supports to help with self-control(the desire for continuity and stability of family support,the need for professionalism and accuracy of healthcare guidance,and the expectation for rationality and matching of resource allocation).Conclusion Healthcare professionals should pay attention to the real experience of COPD patients in the process of disease self-control,help them effectively deal with the challenges of disease self-control,strengthen self-control behaviors,and satisfy their diversified needs by strengthening the support of families,professionals,and the community to improve poor outcomes and reduce the cost of healthcare services.

Objective:To analyze the practical situation of clinical college tutorial system teaching from the perspective of students'needs,and provide reference for optimizing the teaching mode of clinical college tutorial system.Method:A total of 79 undergraduate students who participated in the tutorial system teaching in the clinical college of our university were selected as the research subjects.Through questionnaire surveys and one-to-one in-depth interviews,the current implementation status of the tutorial system teaching mode and students'needs and expectations for the tutorial system teaching mode were understood.Results:The overall satisfaction with the teacher-student relationship was 86.07%,and students'willingness to communicate internally was the main factor affecting communication.And 1.27%of students believed that they had no obstacles in teacher-student communication,and 98.73%of students did not want mentors to teach more than 5 students.And 69.62%of students preferred mentors to provide teaching guidance through clinical practice.Binary logistic regression analysis showed that students'perception that mentorship had no significant effect was a negative factor in teaching effectiveness(OR=0.045,95%CI:0.003-0.598).Conclusions:Taking students'needs as the starting point,strengthening the preaching of the mentorship teaching mode,reinforcing the confidence in learning,and improving the teaching programme and teaching quality evaluation system are conducive to enhancing the quality of mentorship teaching in clinical college.

Periodontitis is a chronic inflammatory disease initiated by biofilm microorganisms and mediated by host immune imbalance.Uncontrolled periodontal infections are the leading cause of tooth loss in adults.Thrombotic diseases can lead to partial or complete obstruction of blood flow in the circulatory system,manifesting as organ or tissue ischemia and necrosis in patients with arterial thrombosis,and local edema,pain and circulatory instability in patients with venous thrombosis,which may lead to mortality or fatality in severe case.Recent studies found that periodontitis might enhance thrombosis through bacterial transmission or systemic inflammation by affecting platelet-immune cell interactions,as well as the coagulation,and periodontal therapy could have a prophylactic effect on patients with thrombotic diseases.In this review,we summarized clinical findings on the association between periodontitis and thrombotic diseases and discussed several novel prothrombotic periodontitis-related agents,and presented a perspective to emphasize the necessity of oral health management for people at high risk of thrombosis.