BACKGROUND:During bone remodeling,bone formation and bone resorption are spatially and temporally coordinated,involving intricate interactions between osteoclasts and osteoblasts.Isoginkgetin,a flavonoid found in Ginkgo biloba,has a wide range of anticancer activity and anti-reactive oxygen species activity;however,the effect of isoginkgetin on osteoclast differentiation is unknown.OBJECTIVE:To study the effect and mechanism of action of isoginkgetin on osteoclastogenesis.METHODS:In vitro studies were performed on mouse bone marrow-derived macrophages,and cell counting kit-8 cytotoxicity assay was used to detect the effect of isoginkgetin on cell viability of bone marrow-derived macrophages.Macrophage colony-stimulating factor and receptor activator of nuclear factor kappa-B ligand were used to induce the differentiation of bone marrow-derived macrophages to osteoclasts.Network pharmacology and molecular docking and molecular dynamics simulations were used to predict the processes and targets of the effects of isoginkgetin on the differentiation of osteoclasts.Tartrate-resistant acid phosphatase staining and F-actin staining were used to detect the effects of isoginkgetin on the differentiation and function of osteoclasts.Western blot and RT-PCR were used to detect the effects of isoginkgetin on the expression of genes and proteins related to osteoclast differentiation,reactive oxygen species,and PI3K/AKT pathways.Fluorescent probes were used to detect cellular and mitochondrial reactive oxygen species levels.Flow cytometry technology was used to detect reactive oxygen species levels in cells.RESULTS AND CONCLUSION:(1)Network pharmacology results showed that isoginkgetin affected osteoporosis mainly through the PI3K-AKT pathway and cellular response to drugs and hypoxia,and GSK3β,ESR1,MCL1 and CCNA2 were the key targets.(2)Cell counting kit-8 and tartrate-resistant acid phosphatase staining results showed that isoginkgetin at 8 μmol/L had the most significant inhibitory effect on osteoclastogenesis in vitro,and F-actin results showed that isoginkgetin inhibited osteoclast cytoskeletal actin ring formation in a concentration-dependent manner.(3)Molecular dynamics simulations showed that isoginkgetin bound well to osteoclastogenesis marker proteins(NFATc1,c-Fos,CTSK,and MMP9).Western blot and RT-PCR results indicated that isoginkgetin inhibited the expression of osteoclastogenesis marker proteins and genes(NFATc1,c-Fos,CTSK,and MMP9).(4)Western blot results showed that isoginkgetin inhibited the phosphorylation level of PI3K/AKT/GSK3β and suppressed osteoclastogenesis by activating the PI3K-AKT-GSK3β pathway.(5)The results of reactive oxygen species assay showed that isoginkgetin significantly reduced receptor activator of nuclear factor kappa-B ligand-induced cellular and mitochondrial reactive oxygen species production,and inhibited the differentiation of bone marrow-derived macrophages to osteoclasts.

BACKGROUND:During bone remodeling,bone formation and bone resorption are spatially and temporally coordinated,involving intricate interactions between osteoclasts and osteoblasts.Isoginkgetin,a flavonoid found in Ginkgo biloba,has a wide range of anticancer activity and anti-reactive oxygen species activity;however,the effect of isoginkgetin on osteoclast differentiation is unknown.OBJECTIVE:To study the effect and mechanism of action of isoginkgetin on osteoclastogenesis.METHODS:In vitro studies were performed on mouse bone marrow-derived macrophages,and cell counting kit-8 cytotoxicity assay was used to detect the effect of isoginkgetin on cell viability of bone marrow-derived macrophages.Macrophage colony-stimulating factor and receptor activator of nuclear factor kappa-B ligand were used to induce the differentiation of bone marrow-derived macrophages to osteoclasts.Network pharmacology and molecular docking and molecular dynamics simulations were used to predict the processes and targets of the effects of isoginkgetin on the differentiation of osteoclasts.Tartrate-resistant acid phosphatase staining and F-actin staining were used to detect the effects of isoginkgetin on the differentiation and function of osteoclasts.Western blot and RT-PCR were used to detect the effects of isoginkgetin on the expression of genes and proteins related to osteoclast differentiation,reactive oxygen species,and PI3K/AKT pathways.Fluorescent probes were used to detect cellular and mitochondrial reactive oxygen species levels.Flow cytometry technology was used to detect reactive oxygen species levels in cells.RESULTS AND CONCLUSION:(1)Network pharmacology results showed that isoginkgetin affected osteoporosis mainly through the PI3K-AKT pathway and cellular response to drugs and hypoxia,and GSK3β,ESR1,MCL1 and CCNA2 were the key targets.(2)Cell counting kit-8 and tartrate-resistant acid phosphatase staining results showed that isoginkgetin at 8 μmol/L had the most significant inhibitory effect on osteoclastogenesis in vitro,and F-actin results showed that isoginkgetin inhibited osteoclast cytoskeletal actin ring formation in a concentration-dependent manner.(3)Molecular dynamics simulations showed that isoginkgetin bound well to osteoclastogenesis marker proteins(NFATc1,c-Fos,CTSK,and MMP9).Western blot and RT-PCR results indicated that isoginkgetin inhibited the expression of osteoclastogenesis marker proteins and genes(NFATc1,c-Fos,CTSK,and MMP9).(4)Western blot results showed that isoginkgetin inhibited the phosphorylation level of PI3K/AKT/GSK3β and suppressed osteoclastogenesis by activating the PI3K-AKT-GSK3β pathway.(5)The results of reactive oxygen species assay showed that isoginkgetin significantly reduced receptor activator of nuclear factor kappa-B ligand-induced cellular and mitochondrial reactive oxygen species production,and inhibited the differentiation of bone marrow-derived macrophages to osteoclasts.

Animal experiments are essential tools in biomedical research, serving as a bridge between basic research and clinical trials. Systematic reviews and meta-analyses (SRs/MAs) of animal experiments are crucial methods for integrating evidence from animal experiment, which can facilitate the translation of findings into clinical research, reduce translational risks, and promote resource integration in basic research. With the continuous development of the Grading of Recommendations, Assessment, Development, and Evaluation (GRADE) methodology, its application in SRs/MAs of animal experiments has gained increasing attention. This article first outlines the principles and specific applications of the GRADE methodology in SRs/MAs of animal experiments, including qualitative descriptive systematic reviews, meta-analyses, and network meta-analyses. It then deeply analyzes the misuse of the GRADE methodology in practice, including incorrect evidence grading, improper classification of evidence, misapplication in qualitative systematic reviews, inconsistencies between the documentation of the upgrading and downgrading process and results, and inappropriate use for making recommendations. Furthermore, this article comprehensively discusses the factors influencing the grading of evidence certainty in SRs/MAs of animal experiments, including the impact of bias risk, indirectness, inconsistency, imprecision, and publication bias on evidence downgrading, as well as the role of large effect sizes and cross-species consistency in evidence upgrading. Finally, in response to the issues discussed, improvement strategies are proposed, including further research and optimization of the GRADE methodology for SRs/MAs of animal experiments, the development of reporting guidelines tailored to the characteristics of SRs/MAs in animal experiment research, and enhanced professional training for researchers in the GRADE methodology. This article aims to improve the quality of evidence in SRs/MAs of animal experiments, strengthen their reliability in clinical decision-making, and promote the more efficient translation of findings from animal experiment research into clinical practice.

Objective To explore the effect of dexmedetomidine nasal spray(DNS)on perioperative sleep quality,anxiety stress in patients undergoing hysterectomy.Methods A total of 75 patients who underwent laparoscopic hysterectomy at Taizhou First People's Hospital from May to December 2024 were selected and divided into control group(37 cases)and experimental group(38 cases)according to the random number table method.Patients in experimental group were respectively sprayed with 50μg or 75pg of DNS at 20:30 on the night before surgery and the night after surgery,patients in control group were respectively given an equal volume of normal saline nasal spray at 20:30 on the night before surgery and the night after surgery.Athens insomnia scale(AIS)scores and sleep quality on the first night of admission(T0),the first night before surgery(T1),the first night after surgery(T2),and the second night after surgery(T3),the scores of hospital anxiety and depression scale(HADS),salivary cortisol andα-amylase on the first day of admission,before entering the operating room,and on the first and second days after surgery,and incidence of adverse reactions 60 minutes after administration between two groups of patients were observed and compared.Results Compared with control group,the AIS scores in experimental group at T1,T2 and T3 were significantly lower,the latency to sleep was significantly shortened,the total sleep time and non-rapid eye movement sleep time were significantly prolonged,and rapid eye movement sleep time was significantly shortened(P＜0.05),number of awakenings at T1 and T2 decreased significantly(P＜0.05),and HADS score,salivary cortisol and α-amylase levels before entering the operating room,and on the first and second days after surgery were all significantly decreased(P＜0.05).There was no statistically significant difference in the incidence of adverse reactions within 60 minutes after administration between two groups of patients(P＞0.05).Conclusion Perioperative administration of DNS can significantly improve the sleep quality of patients undergoing laparoscopic hysterectomy,relieve anxiety,reduce stress levels,and has high safety.

Objective To explore the effect of dexmedetomidine nasal spray(DNS)on perioperative sleep quality,anxiety stress in patients undergoing hysterectomy.Methods A total of 75 patients who underwent laparoscopic hysterectomy at Taizhou First People's Hospital from May to December 2024 were selected and divided into control group(37 cases)and experimental group(38 cases)according to the random number table method.Patients in experimental group were respectively sprayed with 50μg or 75pg of DNS at 20:30 on the night before surgery and the night after surgery,patients in control group were respectively given an equal volume of normal saline nasal spray at 20:30 on the night before surgery and the night after surgery.Athens insomnia scale(AIS)scores and sleep quality on the first night of admission(T0),the first night before surgery(T1),the first night after surgery(T2),and the second night after surgery(T3),the scores of hospital anxiety and depression scale(HADS),salivary cortisol andα-amylase on the first day of admission,before entering the operating room,and on the first and second days after surgery,and incidence of adverse reactions 60 minutes after administration between two groups of patients were observed and compared.Results Compared with control group,the AIS scores in experimental group at T1,T2 and T3 were significantly lower,the latency to sleep was significantly shortened,the total sleep time and non-rapid eye movement sleep time were significantly prolonged,and rapid eye movement sleep time was significantly shortened(P＜0.05),number of awakenings at T1 and T2 decreased significantly(P＜0.05),and HADS score,salivary cortisol and α-amylase levels before entering the operating room,and on the first and second days after surgery were all significantly decreased(P＜0.05).There was no statistically significant difference in the incidence of adverse reactions within 60 minutes after administration between two groups of patients(P＞0.05).Conclusion Perioperative administration of DNS can significantly improve the sleep quality of patients undergoing laparoscopic hysterectomy,relieve anxiety,reduce stress levels,and has high safety.

Objective:To investigate the effect of the magnetic stimulation of the sacral nerve on detrusor weakness after spinal cord injury. Method:36 individuals with detrusor weakness after spinal cord injury were divided into experimental group and control group after randomization.Both groups received conventional rehabilitation treatment,besides,the experimental group received repetitive functional magnetic stimulation(rFMS)on S3 nerve roots.Before treat-ment and after 4-weeks treatment,the simple bladder capacity,bladder ultrasound,self-rating anxiety scale were observed. Result:The maximum automatic micturition volume,average automatic micturition volume,residual urine vol-ume,initial uriesthesis,and anxiety self-evaluation score of the two groups were improved after intervention.And the improvements are more in experimental group(P＜0.05). Conclusion:The effect of the rFMS on individuals with detrusor weakness after spinal cord injury is signifi-cantly better than conventional intervention,which provides a new idea for the clinical treatment of neurogenic bladder with detrusor weakness.

Objective To develop nurses'knowledge,attitude and practice questionnaire on medication management for patients with dysphagia,and test its reliability and validity.Methods Based on the evidence-based summary of the best evidence of medication management for patients with dysphagia,guided by the the-ory of knowledge,attitude and practice,the basic dimensions and item pool of the questionnaire were deter-mined through group discussion,Delphi expert consultation and pre-investigation.In order to revise the ques-tionnaire,437 nurses from 10 tertiary hospitals in Jiangsu Province were conveniently selected for investigation,and the reliability and validity of the questionnaire were tested according to the survey results.Results The nurses'knowl-edge,attitude and practice questionnaire on medication management for patients with dysphagia included 43 items in three dimensions.The three dimensions were analyzed by exploratory factors,and six common factors with characteristic roots>1 were extracted.Two factors were extracted from the knowledge dimension,and the cumulative variance contribution rate was 74.958%,One factor was extracted from the attitude dimen-sion,and the cumulative variance contribution rate was 77.655%.Three factors were extracted from the prac-tice dimension,and the cumulative variance contribution rate was 72.274%.The factor load of each item was 0.618-0.902,Cronbach's α coefficient of the total questionnaire was 0.949,and the test-retest reliability was 0.909.The overall content validity coefficient of the questionnaire was 0.922,and the content validity coeffi-cient for each item was 0.800-1.000.Conclusion The nurses'knowledge,attitude and practice questionnaire on medication management for patients with dysphagia developed in this study has good reliability and validi-ty,and could be used as an effective tool to evaluate the status quo of nurses'medication management for pa-tients with dysphagia.

Objective To explore the clinical features,diagnostic methods,and treatment strategies for severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)encephalitis confirmed through cerebrospinal fluid(CSF)analysis.Methods The clinical data of patients diagnosed with SARS-CoV-2 encephalitis through CSF analysis in the Neurology Intensive Care Unit of the First Affiliated Hospital of Nanchang University from March 2022 to March 2023 were collected.Additionally,the relevant literature published in both domestic and international databases was analyzed and synthesized.Results The main neurological manifestations of five cases included decreased consciousness(5/5),psychiatric disorder(2/5),seizures(2/5),quadriplegia(1/5),and headaches(1/5).Two cases had abnormal brain magnetic resonance imaging(MRI)changes,involving the temporal lobe,insular lobe,thalamus,hippocampus,and pons.Additionally,CSF analysis showed mildly elevated protein levels in two cases.Next-generation sequencing(NGS)of the CSF identified SARS-CoV-2 in all five cases(sequence:41-1620),and human herpesvirus 1 in one case(sequence:21).The treatment regimen for all cases included antiviral therapy,three were additionally treated with glucocorticoids and one received immunoglobulin therapy.All cases achieved a favorable outcome(mRS:0-2).Conclusion SARS-CoV-2 has the potential to induce encephalitis/meningitis due to its neurotropic nature.The consideration of this condition is warranted in patients with relevant epidemiological history and symptoms related to the central nervous system.CSF NGS serves as a valuable tool for early diagnosis,while active antiviral therapy and immunotherapy may improve patient outcomes.

Objective:To explore the efficacy and safety of TiRobot combined with three-dimensional imaging in the minimally invasive surgery for pelvic fractures.Methods:A retrospective analysis was conducted of the 40 patients with pelvic fracture who had been treated by fixation with S1 and S2 sacroiliac screws at Department of Orthopaedics and Traumatology, Yangjiang People's Hospital from January 2019 to May 2021. They were divided into 2 groups according to their treatment methods. In the TiRobot group of 20 cases subjected to percutaneous sacroiliac screw fixation assisted by TiRobot combined with three-dimensional imaging, there were 13 males and 7 females with an age of (38.2 ± 8.8) years. In the manual group of 20 cases subjected to fixation with manual placement of sacroiliac screws under conventional C-arm fluoroscopy, there were 11 males and 9 females with an age of (37.3 ± 9.2) years. The 2 groups were compared in terms of fluoroscopy time for screw placement, guide needle adjustment, operation time, intraoperative blood loss, visual analogue scale (VAS) 72 hours after operation, postoperative hospital stay, time to ambulation, excellent to good rate of screw placement, complication rate, fracture union time, Majeed score at 6 months after operation, and excellent to good rate of functional evaluation.Results:There was no significant difference between the 2 groups in their preoperative general data, showing they were comparable ( P > 0.05). In the TiRobot group, fluoroscopy time for screw placement [(8.2 ± 2.9) s], guide needle adjustment [(0.4 ± 0.2) times], operation time [(67.4 ± 5.5) min], and intraoperative blood loss [(36.5 ± 8.0) mL] were significantly less than those in the manual group [(40.4 ± 4.5) s, (8.6 ± 0.7) times, (78.4 ± 7.2) min, and (41.6 ± 7.8) mL], postoperative VAS [3.0 (4.0, 5.0) points] was significantly lower than that in the manual group [4.0 (5.0, 6.0) points], the excellent to good rate of screw placement (100%, 40/40) was significantly higher than that in the manual group (85.0%, 34/40), and the complication rate (5.0%,1/20) was significantly lower than that in the manual group (35.0%, 7/20) (all P < 0.05). There was no significant difference between the 2 groups in postoperative hospital stay, time to ambulation, fracture union time, Majeed score, or excellent to good rate of functional evaluation ( P > 0.05). Conclusion:In the minimally invasive surgery for pelvic fractures, TiRobot combined with three-dimensional imaging leads to positive outcomes, because it can reduce operation time and radiation exposure, improve accuracy of screw placement, and increase safety.

To effectively solve the serious impact of high oil in the kitchen wastewater on the downstream treatment process, an excellent oil-degrading strain Aeromonas allosaccarophila CY-01 was immobilized to prepare Chitosan-Aeromonas pellets (CH-CY01) by using chitosan as a carrier. Oil degradation condition and efficiency of CH-CY01 pellets were assessed. The growth of immobilized CH-CY01 was almost unaffected, and the maximum degradation rate of soybean oil was 89.7%. Especially at 0.5% NaCl concentration, oil degradation efficiency of CH-CY01 was increased by 20% compared with free cells. In the presence of a surfactant (sodium dodecylbenzene sulfonate) at 1 mg/L, the degradation efficiency of oil by CH-CY01 was increased by 40%. Moreover, using the high-oil catering wastewater as the substrate, more than 80% of the solid oil was degraded with 1% (V/V) CH-CY01 pellets treatment for 7 days, significantly higher than that of free cells. In summary, immobilized CH-CY01 significantly improved the efficiency of oil degradation.
Aeromonas ; Chitosan ; Surface-Active Agents ; Waste Water