Diabetic retinopathy(DR), a major microvascular complication of diabetes, is driven by hyperglycemia-induced oxidative stress, chronic inflammation, and neurodegeneration. Post-translational modifications(PTM)play pivotal roles in DR progression by dynamically regulating protein functions. Key PTMs, including phosphorylation, acetylation, ubiquitination, and O-glcNAcylation, collectively exacerbate vascular dysfunction, inflammatory responses, metabolic dysregulation, and neuronal damage. The intricate crosstalk among PTM underscores the multifaceted pathology of DR. Future research should focus on elucidating PTM interaction networks, developing targeted modulators, and leveraging advanced technologies to uncover their roles in retinal cellular heterogeneity, thereby advancing precision therapeutic strategies for DR.

AIM:This study aims to investigate the mechanisms through which Astragaloside IV(AS)pre-vents and treats osteoporosis by regulating intestinal flora.METHODS:Thirty 3-month-old female Sprague-Dawley(SD)rats were selected for the study.Ten rats were randomly assigned to a sham group,while the remaining twenty underwent bilateral ovariectomy(OVX)to simulate osteoporosis.Following the modeling,the twenty OVX rats were randomly divid-ed into two groups:the OVX group and the AS treatment group,which received continuous gavage for 12 weeks.Bone mineral density(BMD)of the femur and lumbar vertebrae was measured using dual-energy X-ray absorptiometry(DXA).Hematoxylin-eosin(HE)staining was employed to assess the microstructure of the femur and colonic mucosa,while immu-nohistochemistry was used to measure the expression of collagen type Ⅰ alpha 1 chain(COL1A1)protein in the femur.Ad-ditionally,RT-qPCR was utilized to analyze the mRNA expression of bone formation-related indicators,including alkaline phosphatase(ALP),COL1A1,and Runt-related transcription factor 2(RUNX2).Fresh fecal samples were collected from the rats for 16S rDNA sequencing to detect changes in intestinal microbiota composition.RESULTS:Compared to the sham group,OVX rats exhibited a significant increase in body weight and a marked decrease in femur and lumbar ver-tebrae bone density.HE staining revealed trabecular bone fractures with a disrupted reticular structure in the OVX group,along with the presence of numerous cavities and fat vacuoles in the bone marrow.The colonic mucosa showed signs of vil-lous shedding and mild crypt atrophy.Immunohistochemistry results demonstrated a substantial reduction in brown-yellow granules and COL1A1 expression in the OVX group.Conversely,in the AS group,there was a reduction in body weight and a significant increase in bone density of the femur and lumbar vertebrae.The trabecular architecture appeared more or-ganized,with less severe fractures compared to the OVX group.In the AS group,the number of cavities and fat vacuoles in the bone marrow was also reduced,and the colonic mucosa exhibited improved villous structure and less crypt atrophy.Immunohistochemical analysis indicated that AS treatment significantly enhanced COL1A1 expression.Furthermore,after AS intervention,the mRNA expression levels of ALP,COL1A1,and RUNX2 were notably increased.16S rDNA sequenc-ing revealed a significant increase in the abundance of Firmicutes,Proteobacteria,f_Pseudonocardiaceae,f_Marinifilace-ae,f_Oscillospiraceae,f_Ruminococcaceae,and f_Peptostreptococcaceae,while p_Euryarchaeota,Bacteroidetes,and f_Muribaculaceae showed significant reductions.Overall,OVX led to increased diversity in the species distribution of in-testinal microbiota,whereas AS treatment helped recalibrate the aforementioned phyla(families)and reduce diversity.CONCLUSION:Astragaloside IV can increase bone density in OVX rats,improve bone microstructure,promote bone formation,and prevent colonic mucosal damage by regulating the relative abundance of intestinal flora.

AIM:This study aims to investigate the mechanisms through which Astragaloside IV(AS)pre-vents and treats osteoporosis by regulating intestinal flora.METHODS:Thirty 3-month-old female Sprague-Dawley(SD)rats were selected for the study.Ten rats were randomly assigned to a sham group,while the remaining twenty underwent bilateral ovariectomy(OVX)to simulate osteoporosis.Following the modeling,the twenty OVX rats were randomly divid-ed into two groups:the OVX group and the AS treatment group,which received continuous gavage for 12 weeks.Bone mineral density(BMD)of the femur and lumbar vertebrae was measured using dual-energy X-ray absorptiometry(DXA).Hematoxylin-eosin(HE)staining was employed to assess the microstructure of the femur and colonic mucosa,while immu-nohistochemistry was used to measure the expression of collagen type Ⅰ alpha 1 chain(COL1A1)protein in the femur.Ad-ditionally,RT-qPCR was utilized to analyze the mRNA expression of bone formation-related indicators,including alkaline phosphatase(ALP),COL1A1,and Runt-related transcription factor 2(RUNX2).Fresh fecal samples were collected from the rats for 16S rDNA sequencing to detect changes in intestinal microbiota composition.RESULTS:Compared to the sham group,OVX rats exhibited a significant increase in body weight and a marked decrease in femur and lumbar ver-tebrae bone density.HE staining revealed trabecular bone fractures with a disrupted reticular structure in the OVX group,along with the presence of numerous cavities and fat vacuoles in the bone marrow.The colonic mucosa showed signs of vil-lous shedding and mild crypt atrophy.Immunohistochemistry results demonstrated a substantial reduction in brown-yellow granules and COL1A1 expression in the OVX group.Conversely,in the AS group,there was a reduction in body weight and a significant increase in bone density of the femur and lumbar vertebrae.The trabecular architecture appeared more or-ganized,with less severe fractures compared to the OVX group.In the AS group,the number of cavities and fat vacuoles in the bone marrow was also reduced,and the colonic mucosa exhibited improved villous structure and less crypt atrophy.Immunohistochemical analysis indicated that AS treatment significantly enhanced COL1A1 expression.Furthermore,after AS intervention,the mRNA expression levels of ALP,COL1A1,and RUNX2 were notably increased.16S rDNA sequenc-ing revealed a significant increase in the abundance of Firmicutes,Proteobacteria,f_Pseudonocardiaceae,f_Marinifilace-ae,f_Oscillospiraceae,f_Ruminococcaceae,and f_Peptostreptococcaceae,while p_Euryarchaeota,Bacteroidetes,and f_Muribaculaceae showed significant reductions.Overall,OVX led to increased diversity in the species distribution of in-testinal microbiota,whereas AS treatment helped recalibrate the aforementioned phyla(families)and reduce diversity.CONCLUSION:Astragaloside IV can increase bone density in OVX rats,improve bone microstructure,promote bone formation,and prevent colonic mucosal damage by regulating the relative abundance of intestinal flora.

Objective To explore the characteristics of fluid flow within loaded osteons under different boundary conditions.Methods The COMSOL Multiphysics software was used to establish a three-dimensional(3D)finite element model of osteons with different boundary conditions,and the variation rules of pore pressure and flow velocity of osteons under different inner wall pulsating blood pressures and outer wall elastic constraint conditions were analyzed.Results As the pulsatile blood pressure inside the osteon increased from 0 mmHg(1 mmHg=0.133 kPa)to 300 mmHg,the peak pore pressure within the osteon correspondingly increased from 26 kPa to 68 kPa.As the elastic constraint on the outer wall of osteons changed from being completely elastic to completely constrained,the peak pore pressure within osteons increased from 15 kPa to 26 kPa,and the peak flow velocity increased from 0.04 um/s to 0.07 um/s.Conclusions This study reveals the influence laws of changes in boundary conditions such as the pulsatile blood pressure on the inner wall and the elastic constraint on the outer wall of osteons on fluid flow characteristics within loaded osteons.These findings are conducive to a deeper understanding of the mechanical response mechanisms of bone tissues in both physiological and pathological states,and provide an important theoretical basrs for further researches on bone mechanotransduction.

Objective To explore the characteristics of fluid flow within loaded osteons under different boundary conditions.Methods The COMSOL Multiphysics software was used to establish a three-dimensional(3D)finite element model of osteons with different boundary conditions,and the variation rules of pore pressure and flow velocity of osteons under different inner wall pulsating blood pressures and outer wall elastic constraint conditions were analyzed.Results As the pulsatile blood pressure inside the osteon increased from 0 mmHg(1 mmHg=0.133 kPa)to 300 mmHg,the peak pore pressure within the osteon correspondingly increased from 26 kPa to 68 kPa.As the elastic constraint on the outer wall of osteons changed from being completely elastic to completely constrained,the peak pore pressure within osteons increased from 15 kPa to 26 kPa,and the peak flow velocity increased from 0.04 um/s to 0.07 um/s.Conclusions This study reveals the influence laws of changes in boundary conditions such as the pulsatile blood pressure on the inner wall and the elastic constraint on the outer wall of osteons on fluid flow characteristics within loaded osteons.These findings are conducive to a deeper understanding of the mechanical response mechanisms of bone tissues in both physiological and pathological states,and provide an important theoretical basrs for further researches on bone mechanotransduction.

Objective:To determine the relationship between tibial plateau stresses and malunion by exploring the changes in mechanical conduction in the knee joint after malunion of Hoffa fracture of the tibial plateau.Methods:This study selected 28 knee joint specimens treated with formalin for preservation, half of which were from male and half from female individuals with an age of (51.4±9.5) years. Their structures were intact, and flexion-extension activities normal. X-ray examinations excluded osteoporosis, tuberculosis, and diseases that could have potentially affected bone quality. The knee specimens were divided into a control group (intact tibia) ( n=4) and 6 groups of tibial plateau Hoffa fracture malunion model: 3 vertical malunion groups (groups V1, V2, and V3, with a vertical displacement of 1, 2, and 3 mm, respectively, n=4) and 3 separation malunion groups (groups S3, S5, and S7, with a separation displacement of 3, 5, and 7 mm, respectively), with half males and half females in each group. After a 600N vertical load was applied at passive knee flexions at 0°, 30°, 60°, 90°, and 120°, the stress levels in the medial and lateral compartments of the knee joint were measured using pressure-sensitive films. Results:Under a vertical load of 600 N, when the knee joint was in a neutral position (flexion of 0°), the differences in the medial and lateral tibial plateau stress values were not statistically significant between the malunion models groups and the control group ( P>0.05). When the knee flexion increased to 30°, the medial tibial plateau stress in the V3 and S7 groups was significantly greater than that in the control group ( P<0.05). At a knee flexion of 60°, the medial plateau stress was significantly greater in the V3, S5 and S7 groups than that in the control group, and the differences were significantly greater than the comparisons at a knee flexion of 30° (all P<0.05). When the knee flexion was 90°, the medial plateau stress in the V2, V3, S5 and S7 groups was significantly greater than that in the control group ( P<0.05), but the lateral tibial plateau stress in the V3 group was significantly smaller than that in the control group ( P<0.05). When the knee flexion was further increased to 120°, the differences in the medial and lateral plateau stress values were statistically significant between all the malunion groups and the control group ( P<0.05), and the differences significantly greater than the comparisons at a knee flexion of 90° (all P<0.05). Under a vertical load of 600 N, the differences in the stresses on the medial and lateral plateaus were not statistically significant between the control group and all the malunion groups at a knee flexion of 0° ( P>0.05). When the knee flexion increased to 30°, the difference between the medial and lateral stresses was not statistically significant in the control group ( P>0.05), but was statistically significant in the V3 and S7 groups ( P<0.05). When the knee flexion reached 60°, 90°, and 120°, the differences between the medial and lateral tibial plateau stresses in all the groups were statistically significant ( P<0.05). Conclusions:The peak knee stresses after malunion of Hoffa fracture of the tibial plateau correlate with the severity of malunion and knee flexion angles. The mechanical properties are not significantly different between a mild malunion knee and a normal knee, but a significant displacement (vertical displacement >2 mm and separation displacement ≥5 mm) may increase the peak knee stresses to increase the risk of knee osteoarthritis. When the severity of malunion is certain, an increase in knee flexion angle increases the difference in the peak stress between the medial and lateral tibial plateaus, thus increasing the risk of knee osteoarthritis.

OBJECTIVE To investigate the intervention effect of kushenol F （KSC-F） on ulcerative colitis （UC） mice. METHODS Totally 30 male C57BL/6J mice were randomly divided into the normal group， model group， positive drug group （sulfasalazine， 703 mg/kg）， KSC-F 50 mg/kg group （KSC-F50 group）， and KSC-F 100 mg/kg group （KSC-F100 group）， with 6 mice in each group. Except for the normal group， the mice in the remaining groups were given 3% dextran sulfate sodium solution continuously for 7 days to induce UC model. Concurrently， administration groups received corresponding drug solution intragastrically， once a day， for 10 consecutive days. During the experiment， the changes in body weight and bowel movements of the mice were observed. Disease activity index scoring was performed after the last administration. The histopathological morphology of colonic tissue was examined. The levels of inflammatory factors in the serum and colon tissue were measured. Additionally， the mRNA expression of inflammatory factors， and the protein expressions of inflammation-related proteins [interleukin-1β （IL-1β）， forkhead box O1（FOXO1）， phosphoinositide 3-kinase（PI3K）， phosphorylated PI3K（p-PI3K）， p38 mitogen-activated protein kinase（p38 MAPK）， phosphorylated p38 MAPK（p-p38 MPAK） and phosphorylated protein kinase B（p- Akt）] were determined in colonic tissue. RESULTS KSC-F could alleviate weight loss and colonic tissue damage in UC mice. KSC- F reduced the levels of IL-1β， IL-6， IL-8 and tumor necrosis factor-α （TNF-α） in serum， as well as IL-1β， IL-6， IL-17 and TNF- α in colonic tissue to varying degrees and increased the levels of IL-10 in both serum and colonic tissue （P＜0.05 or P＜0.01）. Moreover， KSC-F decreased the expression levels of IL-1β， IL-17 and TNF-α mRNA， as well as p-PI3K， p-p38 MAPK， and p- Akt proteins in colonic tissue to varying degrees， and increased the expression levels of IL-10 mRNA and FOXO1 protein in colonic tissue （P＜0.05 or P＜0.01）. CONCLUSIONS KSC-F effectively alleviates UC symptoms in mice by inhibiting PI3K， Akt and p38 MAPK activation， mitigating the release of pro-inflammatory factors such as IL-1β， IL-6， TNF- α，promoting the anti-inflammatory factor IL-10 secretion， and reducing inflammation-induced colonic tissue damage.

Objective To study the differences and mechanisms of damage to the reproductive organs of male rats by single and compound exposure to microwaves at 2.856 and 9.375 GHz.Methods A total of 40 male Wistar rats were randomly divided into sham group,S10 group,X10 group and SX5 group.Microwavesat 2.856 and 9.375 GHz were used to expose the rats for 6 min in the S10 and X10 groups with an average power density of 10 mW/cm2,respectively.The SX5 group was sequentiallyexposed to 2.856 and 9.375 GHz microwaves with an average power density of 5 mW/cm2 for 6 min.At 1 and 7 d after exposure,the sperm viability and serum sex hormones were detected by light microscopy and electron microscopy,and testicular tissue structure and oxidative stress and energy metabolism levels were examined.Results The sperm viability,testosterone(T),follicle stimulating hormone(FSH),and inhibin B(INHB)decreased in the S10 and X10 groups at 1 and 7 d after exposure(P＜0.01),and in the SX5 group at 7 d after exposure(P＜0.05).The LH decreased in all the exposure groups at 1 d after exposure(P＜0.01),and increased in the S10 and X10 groups at 7 d after exposure(P＜0.05).The spermatogenic epithelium of testicular tissue was lax,spermatogenic cells were edematous and vacuolated,chromatin condensed and shifted side by side,and the damage was significant in the S10 and X10 groups as compared with the SX5 group.The superoxide dismutase(SOD)activity in testis tissue decreased and malondialdehyde(MDA)content increased at 1 and 7 d after exposure in the S10 group(P＜0.01).In the X10 group,the SOD decreased at 1 d after exposure(P＜0.01).The lactate dehydrogenase(LDH)and succinate dehydrogenase(SDH)activity and adenosine triphosphate(ATP)content in testis tissue decreased at 1 and 7 d after exposure in the S10 and X10 groups(P＜0.05).In the SX5 group,the LDH and SDH decreased at 1 d after exposure(P＜0.05).Conclusion Single and combined exposure to S-band and X-band microwaves can cause damage to male reproductive organs.The S-band causes damage more significantly than that of X-band.Single-frequency microwave high-intensity exposure causes damage more significantly than that of multi-frequency microwave prolonged combined exposure.The damage is closely related to oxidative stress and energy metabolism.

ObjectiveTo explore the current status and issues regarding the application of ancient books in clinical practice guidelines and expert consensus of traditional Chinese medicine （TCM） published in China， and to provide methodological recommendations for the incorporation of ancient books in the development of TCM guidelines. MethodsWe searched China National Knowledge Infrastructure （CNKI）， WanFang Data， VIP， SinoMed， PubMed， Embase， as well as six industry websites including China Association of Chinese Medicine， National Group Standards Information Platform， and Chinese Association of the Integration of Traditional and Western Medicine，etc. TCM clinical practice guidelines or expert consensus issued during January 1st， 2017， to November 26th， 2022 were searched. Clinical practice guidelines or expert consensus that explicitly referred to ancient books were included， and the content regarding the searching for ancient books， sources of access to ancient books， methods of evaluating the level of evidence， methods of evaluating the level of recommendation， and methods of evaluating the evidence for the ancient books were analysed. ResultsA total of 1,215 TCM clinical practice guidelines or expert consensus were retrieved， with 442 articles explicitly mentioning the application of ancient books， including 300 （67.87%） clinical practice guidelines and 142 （32.13%） expert consensus. Sixty of the 442 publications explicitly reported that ancient books searching had been conducted （13.57%）； among these 60 publications 27 （45.00%） explicitly reported ancient books searching strategies， and the most frequent method was manual searching with a total of 24 articles （40.00%）. The most popular search source was Chinese Medical Dictionary， a TCM classics database， with a total of 18 articles. 197 articles （44.57%） explicitly reported the evaluation criteria for the level of evidence， of which 141 articles （71.57%） involved the evaluation criteria for the ancient books； 413 articles （93.44%） mentioned ancient books in the recommendations， and only the source of formula name was mentioned in 409 （99.03%） of the publications. ConclusionThe current application of ancient books in TCM clinical practice guidelines and expert consensus is limited， with issues of non-standard searching and evaluation methods. Standar-dization and uniformity are needed in evidence grading and recommendation standards. Future research should clarify the scope and methods of applying ancient book， emphasize their integration with modern research evidence， and enhance their value and quality in the development of TCM clinical practice guidelines.

AIM:This study aims to investigate the effects of Osthole(OST)on inflammation and cartilage-de-grading proteases in an interleukin 1β(IL-1β)-induced chondrocyte inflammation model.METHODS:Prediction:we collected target genes for OST and those related to knee osteoarthritis(KOA)from four databases.After standardizing the target genes obtained from the UniProt database,two overlapping genes were identified using the Venny tool.Additional-ly,protein interaction relationships were obtained from the STRING database,and core target genes were screened and vi-sualized using Cytoscape software.Enrichment analysis for the overlapping genes was performed through Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathways.Finally,the active drug components and target proteins were validated and visualized through computational analysis.Validation:primary rabbit chondrocytes were iso-lated and cultured in vitro.Cell morphology and toluidine blue staining were employed to confirm chondrocyte identity.An inflammatory injury model was induced using IL-1β.The cells were divided into control,model,low-dose,medium-dose,high-dose OST,and celecoxib groups.Chondrocyte viability was assessed using the CCK-8 method.Western blot and im-munofluorescence techniques were utilized to detect the proteins IL-1β,tumor necrosis factor-alpha(TNF-α),matrix me-talloproteinase 13(MMP-13),and a disintegrin and metalloproteinase with thrombospondin motifs(ADAMTS-4).Gene expression levels of IL-1β,IL-6,TNF-α,and MMP-13 were measured via RT-qPCR.RESULTS:A total of 80 potential OST targets were identified for treating KOA,with 10 core target genes(CASP3,TNF,HIF1A,IL-1β,NFKB1,PARP1,NFE2L2,JAK2,MAPK1,and GSK3B)screened.GO and KEGG analyses further elucidated the molecular mechanisms of OST in KOA treatment,highlighting multiple biological processes and signaling pathways involved.Molecular docking simulations confirmed stable binding of OST to key targets TNF and IL-1β within the IL-17 signaling pathway.Chondro-cytes exhibited long spindle and pavement-like shapes.Based on the effects of varying OST concentrations on chondro-cytes,2.5,5,and 10 μmol/L OST were selected for pharmacodynamic testing.Compared to the model group,OST signif-icantly reduced inflammation in the chondrocyte inflammation model and inhibited the expression of cartilage matrix-de-grading enzymes,showing no statistically significant difference from the celecoxib group.CONCLUSION:OST promotes chondrocyte proliferation and differentiation.It effectively inhibits inflammation in the IL-1β-induced chondrocyte model and mitigates chondrocyte injury.The underlying mechanism may involve the degradation of chondroproteoglycans and the protection of the extracellular matrix.