1.Effect of sacroiliac joint ankylosis on outcomes of L5/S1 transforminal lumbar interbody fusion and lumbar sagittal parameters
Yalei WANG ; Xuezhi WANG ; Tao ZHOU ; Xinxin SHEN ; Ding FANG ; Hongliang CHEN
Chinese Journal of Tissue Engineering Research 2026;30(3):634-641
BACKGROUND:The correlation between sacroiliac joint degeneration and lumbar degenerative disease has been analyzed in the literature in the past,but the clinical efficacy and imaging changes after interbody fusion with sacroiliac joint ankylosis in patients with lumbar degenerative disease have not been reported in the literature.OBJECTIVE:To investigate the effect of sacroiliac joint ankylosis on the clinical efficacy and lumbar sagittal regression after L5/S1 single-segment transforminal lumbar interbody fusion in patients with lumbar degenerative disease.METHODS:Thirty-seven patients who underwent L5/S1 segmental transforminal lumbar interbody fusion for lumbar degenerative disease with sacroiliac joint ankylosis between June 2020 and September 2023 in Affiliated Hospital of Xuzhou Medical University were retrospectively analyzed as group A.Thirty-seven patients with lumbar degenerative disease without sacroiliac joint ankylosis who were matched for general information during the same period were selected as controls in group B.Clinical efficacy was assessed using the Oswestry disability index and visual analog scale for lumbar and lower limb pain.The lumbar sagittal parameters included lumbar anterior convexity angle,lumbar partial anterior convexity angle,and lower lumbar anterior convexity angle.Pfirrmann grading was used to assess the degree of preoperative disc degeneration,postoperative endplate damage and screw loosening,and to record the fusion of the operated segments at the final postoperative follow-up visit.RESULTS AND CONCLUSION:(1)There was no statistically significant difference in age,body mass index,bone mineral density,operation time,intraoperative bleeding,preoperative primary diagnosis and postoperative follow-up time between the two groups(P>0.05).(2)The preoperative Pfirrmann grading of lumbar disc degeneration in group A patients(3.4±0.9)was significantly higher than that of group B(3.1±0.6),and the difference was statistically significant(t=2.059,P=0.044).(3)All patients showed significant improvement in postoperative lumbar sagittal parameters compared with preoperative ones(all P<0.05).During the follow-up period,there was a loss of correction in patients in group A.There was no statistical difference in the lumbar anterior convexity angle,lower lumbar anterior convexity angle,and local anterior convexity angle at the last follow-up compared with the preoperative period(P>0.05).The lumbar anterior convexity angle,lower lumbar anterior convexity angle,and local anterior convexity angle in group A were significantly lower than those of group B patients at both preoperative and final follow-up,and the differences were statistically significant(all P<0.05).(4)There was no statistically significant difference in postoperative endplate injury between the two groups(x2=0.181,P=0.670),and screw loosening was significantly higher in group A than in group B,with a statistically significant difference(x2=4.163,P=0.041).(5)At the last follow-up,the incidence of grade 3 fusion and grade 4 fusion was significantly higher in group A than in group B.The difference in the distribution of fusion grades between the two groups was statistically significant(x2=7.848,P=0.031).(6)The Oswestry disability index and lower limb visual analog scale scores at the last follow-up of both groups were significantly improved compared with the preoperative period(P<0.05).The visual analog scale scores for low back pain at 3 months after surgery and at the last follow-up of group A were significantly higher than those of group B(t=2.010,P=0.048;t=2.133,P=0.036).(7)It is concluded that regardless of whether it is accompanied by sacroiliac joint ankylosis or not,lumbar degenerative disease patients who undergo interbody fusion with foramen magnum can achieve good therapeutic effects,but lumbar degenerative disease patients with sacroiliac joint ankylosis who undergo interbody fusion with foramen magnum at the L5/S1 segments have a poorer improvement of low back pain than patients without sacroiliac joint ankylosis after the operation.Furthermore,patients with preoperative sacroiliac ankylosis who underwent L5/S1 segmental transforminal lumbar interbody fusion had a low fusion rate and were prone to loss of correction of the lumbar sagittal position.
2.Effect of Yangjing Tongluo Prescription on Oxidative Damage of Endometrium in Rats with Intrauterine Adhesion Based on Keap1/Nrf2/HO-1 Signaling Pathway
Jiaying CHEN ; Jing ZENG ; Zhaoling YOU ; Yonglian WANG ; Muya LIU ; Fang ZHOU ; Li TANG ; Sainan TIAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):100-108
ObjectiveTo explore the mechanisms of Yangjing Tongluo prescription (YJTL) in the treatment of intrauterine adhesion (IUA) from the perspective of oxidative stress mediated by the Kelch-like ECH-associated protein 1/nuclear factor erythroid 2-related factor 2/heme oxygenase-1 (Keap1/Nrf2/HO-1) signaling pathway. MethodsA total of 48 rats with normal estrous cycles were selected and randomly divided into a normal group (n=8) and a modeling group (n=40). An IUA rat model was established using a dual-injury method combining surgical curettage and infection. Eight rats were randomly selected from the modeling group for a pilot experiment to confirm successful model establishment. After successful modeling, the remaining 32 rats were randomly divided into a model group, a low-dose YJTL group (YJTL-L), a high-dose YJTL group (YJTL-H), and a Progynova group. Rats in the normal and model groups were administered purified water (15 mL·kg-1) by gavage daily, while rats in the YJTL-L, YJTL-H, and Progynova groups received YJTL at doses of 6.43 and 12.86 g·kg-1 and Progynova at 2.06 × 10-4 g·kg-1, respectively, for 14 consecutive days. The general condition, uterine morphology, and uterine index of the rats were monitored. Histopathological changes in uterine tissue were observed using hematoxylin-eosin (HE) staining. Serum levels of reactive oxygen species (ROS) and glutathione peroxidase (GSH-Px) were measured by enzyme-linked immunosorbent assay (ELISA). Protein expression levels of Keap1, Nrf2, and HO-1 in endometrial tissue were detected by Western blot. Immunofluorescence (IF) was used to assess the distribution of Nrf2 and HO-1, as well as the expression of Nrf2 in the cytoplasm and nucleus. ResultsCompared with the normal group, rats in the model group exhibited poor mental status and reduced mobility, markedly edematous and tortuous uterine morphology, decreased gland number, and inflammatory reactions in the endometrium, along with an increased uterine organ index (P<0.05). Serum ROS levels were significantly increased (P<0.05), while serum GSH-Px levels were significantly decreased (P<0.05). In endometrial tissue, Keap1 protein expression was increased (P<0.05), whereas Nrf2 and HO-1 protein expression was decreased. Mild nuclear translocation of Nrf2 was observed, accompanied by increased relative fluorescence intensity of nuclear Nrf2 and decreased relative fluorescence intensity of cytoplasmic HO-1. Compared with the model group, all treatment groups showed varying degrees of improvement in the above symptoms and pathological changes. Serum ROS levels were reduced (P<0.05), serum GSH-Px levels were increased (P<0.05), Keap1 protein expression in endometrial tissue was decreased, and Nrf2 and HO-1 protein expression was increased in a dose-dependent manner (P<0.05). Notably, significant nuclear translocation of Nrf2 was observed, with correspondingly increased relative fluorescence intensity of nuclear Nrf2 and enhanced relative fluorescence intensity of cytoplasmic HO-1. ConclusionYJTL may enhance antioxidant capacity and repair oxidative damage to the endometrial basal layer by regulating the Keap1/Nrf2/HO-1 signaling pathway.
3.Luotong Xianrong Yin Inhibits Pulmonary Injury in Idiopathic Pulmonary Fibrosis Rats by Regulating STAT3/HIF-1α-mediated Glycolysis
Dongna LI ; Hao WANG ; Chaoyi FANG ; Jiepeng WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):118-125
ObjectiveTo investigate the characteristics and potential mechanisms of Luotong Xianrong Yin (LTXRY) in improving lung injury in rats with idiopathic pulmonary fibrosis (IPF) by regulating glycolysis. MethodsForty specific pathogen-free (SPF) Sprague-Dawley (SD) rats were randomly divided into a sham-operated group (10 mL·kg-1), model group (10 mL·kg-1), LTXRY group (15.18 g·kg-1), and nintedanib group (0.1 g·kg-1), with 10 rats in each group. The IPF rat model was established by intratracheal instillation of bleomycin. After 28 days of gavage intervention, pulmonary function was assessed. Lung pathological changes were observed by hematoxylin-eosin (HE) and Masson staining. Enzyme-linked immunosorbent assay (ELISA) was used to determine the levels of inflammatory factors, including tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-6, in lung tissue. Chemiluminescence assays were employed to detect lactate content and lactate dehydrogenase activity in lung tissue. Western blot was used to measure the protein expression of transforming growth factor-β1 (TGF-β1), CollagenⅠ and CollagenⅢ to evaluate collagen deposition, as well as hexokinase 2 (HK2), pyruvate kinase M2 (PKM2), and 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3) to assess glycolysis levels. Network pharmacology was applied to analyze the potential targets and signaling pathways of LTXRY in IPF, and molecular docking was conducted to evaluate the binding energy between active components and potential targets. Western blot was further used to detect the expression of target- and pathway-related proteins. ResultsCompared with the sham-operated group, rats in the model group showed significantly increased main airway resistance (Rn) and respiratory system resistance (Rrs), and significantly decreased respiratory system compliance (Crs). Inflammatory infiltration and collagen deposition were observed in lung tissue, with significantly increased levels of TNF-α, IL-1β, and IL-6, as well as elevated protein expression of TGF-β1, CollagenⅠ and CollagenⅢ. Lactate content, lactate dehydrogenase activity, and the protein expression of HK2, PKM2, and PFKFB3 in lung tissue were significantly increased. Network pharmacology analysis indicated that signal transducer and activator of transcription 3 (STAT3) was a key target of LTXRY in IPF, and hypoxia-inducible factor-1 (HIF-1) was a critical signaling pathway. The expression levels of phosphorylated STAT3 (p-STAT3) and HIF-1α in lung tissue were significantly higher than those in the sham-operated group. Compared with the model group, rats in the LTXRY group showed significantly decreased Rn and Rrs and significantly increased Crs. Lung inflammatory infiltration and collagen deposition were markedly alleviated, with significantly reduced levels of TNF-α, IL-1β, and IL-6, and decreased protein expression of TGF-β1, CollagenⅠ and CollagenⅢ. Lactate content, lactate dehydrogenase activity, and the protein expression of HK2, PKM2, and PFKFB3 were significantly decreased, accompanied by markedly reduced expression of p-STAT3 and HIF-1α. ConclusionLTXRY alleviates lung tissue injury in IPF rats by regulating glycolysis mediated by the STAT3/HIF-1α signaling pathway.
4.Clinical Observation on Prevention of Recurrence of Common Bile Duct Stones After ERCP with Yuyin Lidan Granules
Xiao WANG ; Yong FANG ; Cong HE ; Jiali ZHANG ; Meng YU ; Jing KONG ; Yi JIANG ; Chuanqi CHENG ; Xiaosu WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):159-166
ObjectiveTo observe the clinical efficacy and safety of Yuyin Lidan granules (YYLD) in preventing the recurrence of common bile duct stones (CBDS) in patients with liver and gallbladder dampness-heat syndrome following endoscopic retrograde cholangiopancreatography (ERCP). MethodsThis randomized, parallel, controlled trial enrolled postoperative CBDS-ERCP patients who met the inclusion and exclusion criteria. Sixty-four patients were randomly assigned to an observation group or a control group, with 32 cases in each. Both groups received conventional Western medical treatment after ERCP, while the observation group additionally received YYLD for 8 weeks. The follow-up period lasted for 1 year. The efficacy indicators included bile bilirubin levels, traditional Chinese medicine (TCM) syndrome scores, clinical efficacy rate, pancreatitis and inflammation markers, postoperative liver function, and CBDS recurrence rate at 1-year follow-up, which were used to jointly evaluate the clinical efficacy and safety of both groups. ResultsA total of 56 patients completed the study and were included in the final analysis, i.e., 29 in the observation group and 27 in the control group. Baseline characteristics were comparable between the two groups. Compared with pre-treatment and with the control group after treatment, the bile bilirubin level in the observation group significantly decreased (P<0.05). After treatment, the clinical cure and marked improvement rates were higher in the observation group than in the control group, showing a statistically significant difference in overall clinical efficacy (P<0.05). Compared with pre-treatment, the primary and secondary symptoms in the observation group, as well as the primary symptom and the secondary symptom of nausea and vomiting in the control group (weeks 4 and 8), were significantly reduced (P<0.05). Compared with the control group after treatment, the observation group showed significant reductions in the primary symptom of loose stools/constipation (day 5 and week 4) and in three secondary symptoms, i.e., bitter taste and sticky dry mouth, abdominal distension and poor appetite (throughout the treatment period), and general heaviness and fatigue (day 5 and week 4), with statistical differences (P<0.05). Compared with pre-treatment, both groups showed decreased lipase and urinary amylase levels (P<0.05). However, no significant between-group differences were observed in pancreatitis or inflammation-related indices after treatment. Compared with pre-treatment, all liver function indicators in the observation group and alanine aminotransferase ( ALT ), γ-glutamyl transferase ( γ-GT ), alkaline phosphatase (ALP), and conjugated bilirubin in the control group significantly decreased at weeks 4 and 8 (P<0.05). Compared with the control group after treatment, only serum total bilirubin and unconjugated bilirubin were significantly reduced in the observation group during the treatment period (P<0.05). ConclusionYYLD combined with conventional Western medical treatment can effectively regulate bilirubin metabolism (in bile and serum), improve TCM clinical symptoms, and prevent CBDS recurrence after ERCP in patients with liver and gallbladder dampness-heat syndrome. This regimen is safe and effective and is worthy of further clinical research and promotion.
5.From Gene Expression to Transcriptome-wide Association Study: Development and Comparison of Methodology
Kun FANG ; Guozhuang LI ; Linting WANG ; Qing LI ; Kexin XU ; Lina ZHAO ; Zhihong WU ; Jianguo ZHANG ; Nan WU
Medical Journal of Peking Union Medical College Hospital 2026;17(1):223-229
Over the past two decades, genome-wide association study(GWAS) has identified numerous genetic variants and loci associated with heritable diseases. With the gradual maturation and saturation of GWAS methodologies, transcriptome-wide association study(TWAS) offers a novel perspective by linkinggenetic phenotypes to gene expression levels. By integrating TWAS with other multi-omics analyses, researchers can gain a deeper understanding of heritable diseases. This article provides an overview of recent groundbreaking and representative TWAS methods and tools, analyzes their strengths and limitations, and discusses future trends in TWAS development.
6.Mechanisms of Improving Hippocampal Synaptic Plasticity Through GLP-1/GLP-1r Pathway to Alleviate Anxiety and Depression-like Behaviors in Chronic Restraint Stress Rats by Xiaoyaosan
Hao WANG ; Yanan YAN ; Jiepeng WANG ; Chaoyi FANG ; Fang FANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):34-42
ObjectiveTo observe the effects of Xiaoyaosan on glucagon-like peptide-1 (GLP-1)/GLP-1 receptor (GLP-1r) and protein kinase A (PKA)/cAMP response element binding protein (CREB)/brain-derived neurotrophic factor (BDNF) signaling pathways in the hippocampal CA1 region of rats under chronic restraint stress (CRS),and to explore the mechanism of this formula to alleviate anxiety and depression-like behaviors. Methods40 specific pathogen-free male Sprague-Dawley (SD) rats were randomly divided into normal,model,Xiaoyaosan,and fluoxetine groups,with 10 rats in each group. CRS was used to induce anxiety and depression-like behaviors. The rats in the Xiaoyaosan group were gavaged with aqueous solution of traditional Chinese medicine formula granules (7.36 g·kg-1·d-1),while those in the fluoxetine group were gavaged with aqueous solution of fluoxetine (2 mg·kg-1·d-1). Body weight was measured on days 0,7,14,and 21 of the experiment. On days 0 and 22 of the experiment,the sucrose preference test (SPT),forced swimming test (FST),and open field test (OFT) were performed. The pathological morphology of the hippocampal CA1 region was observed by Nissl staining. The relative mRNA expression of post-synaptic density protein-95 (PSD95) and synapsin (SYP) was detected by reverse transcription quantitative real-time polymerase chain reaction. Immunohistochemistry and Western blot were used to detect expression of proteins in the GLP-1/GLP-1r and PKA/CREB/BDNF pathways in the hippocampal CA1 region. ResultsAfter CRS modeling,compared with the normal group,the rats of the model group had anxiety and depression-like behavioral manifestations,neuronal damage in the hippocampal CA1 region,significantly downregulated expression of synaptic plasticity markers PSD95 and SYP genes (P<0.01),and inhibition of GLP-1/GLP-1r and PKA/CREB/BDNF signaling pathways (P<0.05,P<0.01). Compared with the model group,the Xiaoyaosan group exhibited alleviated anxiety and depression-like behaviors,reduced neuronal damage in the hippocampal CA1 region, significantly increased expression of PSD95 and SYP genes (P<0.01),and the activation of the GLP-1/GLP-1r and PKA/CREB/BDNF signaling pathways (P<0.05,P<0.01). ConclusionXiaoyaosan can alleviate anxiety and depression-like behaviors in CRS rats by improving synaptic plasticity in the hippocampal CA1 region. The mechanisms may be related to the activation of the GLP-1/GLP-1r pathway and its mediated PKA/CREB/BDNF signaling pathway by the formula.
7.Mechanisms of Sini San in Regulation of Gut Microbiota Against Depression and Liver Injury in CUMS Rats
Junling LI ; Yan ZHANG ; Lei WANG ; Fang QI ; Zhenzhen CHEN ; Tianxing CHEN ; Yuhang LIU ; Xueying WANG ; Xianwen TANG ; Yubo LI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):33-40
ObjectiveTo explore the efficacy and mechanisms of Sini San in the treatment of depression and liver injury based on gut microbiota. MethodsThirty-two male Sprague-Dawley (SD) rats were randomly divided into a normal group, model group (M), Sini San group (MS, 2.5 g·kg-1), and fluoxetine group (MF, 2 mg·kg-1). Except for the normal group, rats in the other three groups were subjected to chronic unpredictable mild stress (CUMS). After 8 weeks, the open-field test and sucrose preference test were conducted. Enzyme-linked immunosorbent assay (ELISA) was used to detect serum corticosterone (CORT), adrenocorticotropic hormone (ACTH), corticotropin-releasing factor (CRF), lipopolysaccharide (LPS), Zonulin, interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), γ-aminobutyric acid (GABA) levels in the hippocampus and prefrontal cortex, and brain-derived neurotrophic factor (BDNF) levels in the hippocampus. Real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect hippocampal BDNF mRNA expression. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured using the ultraviolet lactate dehydrogenase method. The ultrastructure of the intestinal epithelium was observed by electron microscopy, and gut microbiota in rat feces were analyzed using 16S rDNA high-throughput sequencing. ResultsCompared with the normal group, the sucrose preference of rats in the model group was significantly reduced (P0.01), whereas it was significantly increased in the Sini San group compared with the model group (P0.05). Compared with the normal group, hippocampal GABA protein levels and BDNF mRNA expression in the model group were significantly decreased (P0.05), and compared with the model group, both were significantly increased in the Sini San group (P0.05, P0.01). Compared with the normal group, serum LPS and Zonulin levels in the model group were significantly increased (P0.05, P0.01), and compared with the model group, Zonulin levels in the Sini San group were significantly decreased (P0.05). No obvious changes were observed in the ultrastructure of the jejunal mucosa among groups. Compared with the normal group, widened and blurred tight junctions, sparse and shortened microvilli, and mitochondrial swelling with cristae disruption in epithelial cells were observed in the ileal and colonic mucosa of the model group, which were markedly improved in the Sini San and fluoxetine groups. The results of 16S rDNA high-throughput sequencing showed that Sini San improved CUMS-induced dysbiosis of Bacteroidetes and Proteobacteria. Correlation analysis indicated that Bacteroidetes and Proteobacteria were significantly correlated with depression-related indicators, liver function, and intestinal mucosal permeability. ConclusionSini San exerts antidepressant and hepatoprotective effects by improving Bacteroidetes and Proteobacteria and inhibiting the increase in intestinal mucosal permeability in CUMS rats.
8.Mechanisms of Sini San in Regulation of Gut Microbiota Against Depression and Liver Injury in CUMS Rats
Junling LI ; Yan ZHANG ; Lei WANG ; Fang QI ; Zhenzhen CHEN ; Tianxing CHEN ; Yuhang LIU ; Xueying WANG ; Xianwen TANG ; Yubo LI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):33-40
ObjectiveTo explore the efficacy and mechanisms of Sini San in the treatment of depression and liver injury based on gut microbiota. MethodsThirty-two male Sprague-Dawley (SD) rats were randomly divided into a normal group, model group (M), Sini San group (MS, 2.5 g·kg-1), and fluoxetine group (MF, 2 mg·kg-1). Except for the normal group, rats in the other three groups were subjected to chronic unpredictable mild stress (CUMS). After 8 weeks, the open-field test and sucrose preference test were conducted. Enzyme-linked immunosorbent assay (ELISA) was used to detect serum corticosterone (CORT), adrenocorticotropic hormone (ACTH), corticotropin-releasing factor (CRF), lipopolysaccharide (LPS), Zonulin, interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), γ-aminobutyric acid (GABA) levels in the hippocampus and prefrontal cortex, and brain-derived neurotrophic factor (BDNF) levels in the hippocampus. Real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect hippocampal BDNF mRNA expression. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured using the ultraviolet lactate dehydrogenase method. The ultrastructure of the intestinal epithelium was observed by electron microscopy, and gut microbiota in rat feces were analyzed using 16S rDNA high-throughput sequencing. ResultsCompared with the normal group, the sucrose preference of rats in the model group was significantly reduced (P<0.01), whereas it was significantly increased in the Sini San group compared with the model group (P<0.05). Compared with the normal group, hippocampal GABA protein levels and BDNF mRNA expression in the model group were significantly decreased (P<0.05), and compared with the model group, both were significantly increased in the Sini San group (P<0.05, P<0.01). Compared with the normal group, serum LPS and Zonulin levels in the model group were significantly increased (P<0.05, P<0.01), and compared with the model group, Zonulin levels in the Sini San group were significantly decreased (P<0.05). No obvious changes were observed in the ultrastructure of the jejunal mucosa among groups. Compared with the normal group, widened and blurred tight junctions, sparse and shortened microvilli, and mitochondrial swelling with cristae disruption in epithelial cells were observed in the ileal and colonic mucosa of the model group, which were markedly improved in the Sini San and fluoxetine groups. The results of 16S rDNA high-throughput sequencing showed that Sini San improved CUMS-induced dysbiosis of Bacteroidetes and Proteobacteria. Correlation analysis indicated that Bacteroidetes and Proteobacteria were significantly correlated with depression-related indicators, liver function, and intestinal mucosal permeability. ConclusionSini San exerts antidepressant and hepatoprotective effects by improving Bacteroidetes and Proteobacteria and inhibiting the increase in intestinal mucosal permeability in CUMS rats.
9.Improvement effects and mechanism of astragaloside Ⅳ on neuroinflammation
Mimi WANG ; Yonggang FENG ; Yun HAN ; Kaixin SHAN ; Fuyu LIU ; Mingsan MIAO ; Xiaoyan FANG
China Pharmacy 2026;37(1):30-35
OBJECTIVE To investigate the improvement effects and mechanism of astragaloside Ⅳ (AS- Ⅳ ) on lipopolysaccharide (LPS)-induced neuroinflammation. METHODS BV2 cells were divided into control group, LPS group, AS-Ⅳ groups at concentrations of 20 and 40 μmol/L, and dexamethasone group (2 μmol/L). Except for control group, neuroinflammation model was established with LPS (1 μg/mL) in other groups after medication. The levels of inflammatory factors [interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and nitric oxide (NO)] in cell supernatant were measured in each group. Mice were randomly divided into normal group, model group, positive control group (Aspirin enteric-coated tablet, 20 mg/kg), AS-Ⅳ low- and high-dose groups (10, 20 mg/kg), with 6 mice in each group. Mice in each group were administered the corresponding drug/normal saline via gavage/intraperitoneal injection, once a day, for 14 consecutive days. Except for normal group, other groups were intraperitoneally injected with LPS (250 μg/kg) 1 hour after daily administration of the drug/normal saline to establish neuroinflammation model. Serum levels of IL-6 and TNF-α were measured 2 h after the last medication; histopathological morphology of cerebral tissue in mice were observed; the co-localization of inducible nitric oxide synthase (iNOS)/ionized calcium binding adapter molecule 1 (Iba1) and CD206/Iba1 in the cerebral cortex region of mice was observed; the expressions of proteins related to the nuclear factor-κB (NF-κB)/mitogen-activated protein kinase (MAPK) signaling pathway in brain tissue of mice were also determined, including NF-κB p65, phosphorylated NF-κB p65(p-NF-κB p65), p38 MAPK, phosphorylated p38 MAPK (p-p38 MAPK), extracellular signal-regulated kinase (ERK), and phosphorylated ERK (p-ERK). RESULTS In the cell experiments, compared with control group, the levels of IL-6, TNF- α and NO in the cell supernatant of the LPS group were increased significantly (P<0.05); compared with LPS group, the levels of IL-6, TNF-α and NO were decreased significantly in the administration groups (P<0.05). In the animal experiments, compared with the normal group, the serum levels of IL-6 and TNF- α, the number of iNOS/Iba1 co-localization positive cells in the cerebral cortex, and the phosphorylation levels of p38 MAPK, NF- κB p65 and ERK proteins in brain tissue were all significantly increased/elevated in model group (P<0.05); the number of CD206/ Iba1 co-localization positive cells in the cerebral cortex region significantly decreased (P<0.05). The neurons in the cerebral cortex and the CA3 region of the hippocampus displayed a disordered arrangement. Compared with model group, above quantitative indexes of mice were all reversed significantly in administration groups (P<0.05); the neuronal cells in the cerebral cortex and the CA3 region of the hippocampus exhibited a relatively orderly arrangement. CONCLUSIONS AS-Ⅳ may inhibit the activation of the NF-κB/MAPK signaling pathway, promote the M2-type polarization of microglia, and thereby suppress neuroinflammatory responses.
10.A study on the preparation of a BGN-loaded thermosensitive adhesive and its performance in barrier membrane fixation
WANG Yuzhu ; GU Junting ; LI Zhiting ; BAI Que ; DANG Gaopeng ; WANG Yifei ; SUN Xiaotang ; NIU Lina ; FANG Ming
Journal of Prevention and Treatment for Stomatological Diseases 2026;34(1):41-53
Objective:
To investigate the barrier membrane fixation performance and enhanced guided bone regeneration (GBR) capability of a thermosensitive adhesive containing bioactive glass nanoparticles in order to provide a novel solution for membrane fixation during GBR procedures.
Methods:
M2NP@BGN (methoxyethyl acrylate-co-N-isopropylacrylamide-co-protocatechuic acid@Bioactive glass nanoparticle), a thermosensitive adhesive, was synthesized via free radical polymerization by compositing methoxyethyl acrylate, N-isopropylacrylamide, and protocatechuic acid into a basic adhesive that was modified with bioactive glass nanoparticle (BGN). The successful fabrication of basic adhesive M2NP was characterized by attenuated total reflection-Fourier transform infrared spectroscopy and nuclear magnetic resonance spectroscopy. The thermosensitive adhesive M2NP@BGN (BGN concentration of 1 mg/mL) was characterized by scanning electron microscopy and a rheometer. By adjusting the BGN concentration (0.1 mg/mL, 0.5 mg/mL, 1 mg/mL, and 2 mg/mL), the adhesive and mechanical strengths were investigated with a universal testing machine. Biocompatibility was evaluated with a cell counting kit-8 assay and hemolysis test to identify the optimal formulation. The optimal material’s extract was co-cultured with mouse bone marrow mesenchymal stem cells, and its osteogenic activity was examined in vitro by quantitative real-time PCR, alkaline phosphatase, and alizarin red S staining. The rat mandibular defect model was established, filled with bone graft, and divided into 3 groups based on membrane fixation method: M2NP@BGN (BGN concentration of 1 mg/mL) fixation group (M2NP@BGN), titanium nail fixation group (Nail), and unfixed control group (Negative). Bone regeneration was analyzed after 8 weeks by micro computed tomography and histological staining.
Results:
M2NP@BGN (BGN concentration of 1 mg/mL) was successfully synthesized and demonstrated rapid gelation under warm, humid conditions. The adhesive with a BGN concentration of 1 mg/mL exhibited the highest adhesive strength (P < 0.001) and significantly enhanced mechanical strength (P < 0.001) under 37℃ wet conditions. All formulations showed excellent biocompatibility, with cell viability > 80% and hemolysis ratio < 5%. M2NP@BGN (BGN concentration of 1 mg/mL) significantly upregulated the expression of Runx2 and Col I (P < 0.001) and enhanced the activity of osteogenic differentiation markers (P < 0.05). In the animal model, the M2NP@BGN group (BGN concentration of 1 mg/mL) achieved significantly higher bone volume fraction and better bone maturity compared to the negative and nail groups (P < 0.05).
Conclusion
M2NP@BGN (BGN concentration of 1 mg/mL) combines excellent wet adhesion with potent osteogenic activity, enhances the bone augmentation efficacy of membranes, and presents a novel fixation strategy with significant clinical translation potential for GBR therapy.


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