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.

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.

Objective:To study the variational trend in disease characteristics of patients with hepatitis B-related primary liver cancer (HBV-HCC) in the past five years.Method:A single-center retrospective cross-sectional analysis was performed to compare patients diagnosed with HBV-HCC from January 2012 to December 2016 (control group) and from January 2017 to December 2021 (observation group). The data of the study variables were extracted from the electronic medical record system of the hospital information system of the Second Hospital of Lanzhou University. The 1:2 propensity score matching was used to adjust potential confounding factors such as gender and age. Multivariate logistic regression analysis was used to study the factors affecting changes in disease characteristics of the HBV-HCC population in the observation group. GraphPad Prism 8.0 software was used to draw forest plots to intuitively display the effect size of the study variables in the logistic regression analysis.The t-test was used to compare normally distributed data between groups. The χ2 test was used for inter-group comparison. Results:A total of 1 717 eligible cases were collected, including 510 in the control group and 1 207 in the observation group. Compared with the control group, the number of newly diagnosed cases in the observation group increased by 2.36 times, and males were still the main onset population (83.3% vs. 82.7%). The median age of onset increased (51.9 vs. 53.5 years, P<0.001). 79.4% of HBV-HCC patients had not received antiviral therapy, and the proportion of HBeAg-negative patients increased (56.4%). The factors affecting HBV-HCC patients included family history of HBV ( OR=1.626, 95% CI: 1.181-2.238), family history of hepatocellular carcinoma ( OR=1.388, 95% CI: 1.013-1.901), hypoviremia ( OR=1.322, 95% CI: 1.046-1.671), abnormal alanine aminotransferase ( OR=1.545, 95% CI: 1.231-1.940), liver fibrosis ( OR=1.478, 95% CI: 1.153-1.894), liver cirrhosis ( OR=1.431, 95% CI: 1.128-1.815), and metabolic-related fatty liver disease ( OR=1.438, 95% CI: 1.116-1.815) after propensity score matching adjustment. The factors affecting HBeAg-positive patients were decreased ( OR=0.390, 95% CI: 0.389-0.617); however, the number of early HBV-HCC diagnoses was increased (12.7% vs. 19.3%, P=0.001). Conclusion:The characteristics of patient disease and occurrence of HBV-HCC are changing over the past five years. The risk of developing hepatocellular carcinoma in middle- to older male patients with chronic hepatitis B is increasing with familial history of HBV and hepatocellular carcinoma, HBeAg negativity, hypoviremia, abnormal alanine aminotransferase, liver fibrosis, cirrhosis, and metabolic-related fatty liver disease.

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.

AIM: To investigate the efficacy of optimal pulse technology(OPT)in the treatment of demodex blepharitis and its influence on ocular surface function.METHODS: A retrospective study was conducted from February 2018 to October 2020. A total of 127 patients(254 eyes)with demodex blepharitis were assigned to the observation group and the control group according to the treatment method. The control group(63 patients, 126 eyes)were given conventional hot compress, eye cleansing and drug therapy. On this basis, the observation group(64 patients, 128 eyes)was treated with OPT. Both groups were given 6wk of continuous treatment. Demodex count, Marx's line scores, meibum character scores, ocular surface disease index(OSDI)scores, non-invasive tear break-up time(NIBUT), non-invasive tear meniscus height(NITMH)and lipid layer thickness(LLT)were compared between the two groups, and safety was evaluated.RESULTS: After 6wk of treatment, demodex count, Marx's line scores, meibum character scores and OSDI scores of the two groups decreased. NIBUT, NITMH and LLT increased. Meanwhile, demodex count, Marx's line scores, meibum character scores and OSDI scores of the observation group were significantly lower than those in the control group. NIBUT, NITMH and LLT were longer/larger than those in the control group(P<0.001). No obvious abnormality of intraocular pressure or conjunctival/corneal injury was observed in either group.CONCLUSION:OPT is effective and safe in the treatment of demodex blepharitis.