Objective To discuss the effects of CeO₂-loaded dendritic mesoporous silica nanoparticles （DS@CeO₂ NPs） on the osteogenic differentiation of mouse jaw bone marrow mesenchymal stem cells （JBMMSCs）， and to clarify its possible mechanism. Methods DS@CeO₂ NPs were synthesized. Transmission electron microscopy （TEM） and dynamic light scattering （DLS） method were used to detect the morphology and particle size of DS@CeO₂ NPs. Enzyme digestion method was used to isolate mouse JBMMSCs. Flow cytometry method was used to identify the cell surface markers on JBMMSCs. The cells were divided into blank control group and different concentrations of DS@CeO2 NPs （25， 50， 100， and 200 mg·L^-1） groups. Cell counting kit-8 （CCK-8） method and Calcein/propidium iodide （PI） staining method were used to detect the proliferation and survival of the cells in various groups. The cells were further divided into control group and different concentrations of DS@CeO₂ NPs osteogenic induction groups （0， 25， 50， and 100 mg·L^-1）. After 7 d of osteogenic induction， alkaline phosphatase （ALP） staining method was used to detect the ALP activities in the cells in various groups. Real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of ALP， Runt-related transcription factor 2 （Runx2）， bone sialoprotein 2 （BSP2）， and osteopontin （OPN） mRNA in the cells in various groups. Western blotting method was used to detect the expression levels of ALP， Runx2， and OPN proteins in the cells in various groups. In addition， osteogenesis group and osteogenesis+DS@CeO₂ NPs group were established. RT-qPCR method and Western blotting method were used to detect the expression levels of β-catenin mRNA and protein in the cells in two groups. Under hydrogen peroxide （H₂O₂） stimulation， the cells were divided into control group and DS@CeO₂ NPs pretreatment group. 2'，7'-dichlorodihydrofluorescein diacetate （DCFH-DA） fluorescence staining method and flow cytometry method were used to detect the levels of reactive oxygen species （ROS） in the cells in two groups. Results The TEM and DLS results showed that DS@CeO₂ NPs exhibited a nearly spherical morphology with an average hydrated particle size of 378.2 nm. The flow cytometry results showed that the expression rates of CD73， CD90， CD105， and CD44 in the cells were all higher than 95%， while CD34 and CD45 were almost not expressed， so the cells could be identified as JBMMSCs. The CCK-8 method results showed that compared with control group， there was no statistically significant difference in the survival rates of the cells in 25， 50， and 100 mg·L^-1 DS@CeO₂ NPs groups （P>0.05）， while the survival rate of the cells in 200 mg·L^-1 DS@CeO₂ NPs group was significantly decreased （P<0.05）. The ALP staining results showed that compared with control group and 0 mg·L^-1 DS@CeO₂ NPs group， the cells in 25 and 50 mg·L^-1 DS@CeO₂ NPs groups exhibited darker ALP staining and larger positive staining areas， and the ALP activities in the cells were increased. The RT-qPCR and Western blotting results showed that compared with 0 mg·L^-1 DS@CeO₂ NPs group， the expression levels of ALP， BSP2， Runx2， and OPN mRNA as well as the expression levels of ALP， Runx2， and OPN proteins in the cells in 25 mg·L^-1 DS@CeO₂ NPs group were significantly increased （P<0.05）. Compared with pure osteogenic group， the expression levels of β-catenin mRNA and protein in the cells in osteogenic+DS@CeO₂ NPs group were significantly increased （P<0.05）. The DCFH-DA staining and flow cytometry results showed that compared with control group， the ROS level in the cells in DS@CeO₂ NPs pretreatment group was decreased. Conclusion DS@CeO₂ NPs show good biocompatibility and can promote the osteogenic differentiation of JBMMSCs； the mechanism may be related to activation of Wnt/β-catenin signaling pathway and antioxidant effect.

Objective To investigate the effects of sodium palmitate （PA）-induced lipotoxicity on mitochondrial function and apoptosis in mouse L929 fibroblasts， and to clarify its associations with mitofusin 2 （MFN2） expression and mitochondria-associated endoplasmic reticulum membrane （MAMs） structure. Methods Mouse L929 fibroblasts were used as the model，which were treated with different concentrations of PA （0.1， 0.2， 0.3， 0.4， 0.5， and 0.6 mmol·L^-1） for 12， 24， 36， and 48 h. Cell counting kit-8 （CCK-8） method was used to detect the survival rates of the cells in various groups after treatment， so as to screen the optimal intervention condition of PA. The cells were then divided into control group and PA group. Flow cytometry method was used to detect the apoptotic rates in the cells in two groups. Oil red O staining method was used to observe the lipid deposition in the cells in two groups. Transmission electron microscopy （TEM） was used to observe the structural characteristics of mitochondria and MAMs in the cells in two groups. Flow cytometry was used to detect the levels of reactive oxygen species （ROS） in the cells in two groups. JC-1 method was used to detect the mitochondrial membrane potential in the cells in two groups. Real-time fluorescence quantitative PCR （RT-qPCR） method and Western blotting method were used to detect the expression levels of MFN2 mRNA and protein in the cells in two groups. Results The CCK-8 assay results showed that at 12， 24， 36， and 48 h post treatment， compared with control group， the survival rates of L929 cells in groups treated with PA at concentrations ≥0.2 mmol·L^-1 were significantly decreased （P<0.05 or P<0.001）， showing a concentration- and time-dependent effect. The survival rate of L929 cells was approximately 60% in 0.3 mmol·L^-1 PA group after 24 h of treatment， which was used as the subsequent PA intervention condition. The flow cytometry and Oil red O staining results showed that compared with control group， the apoptotic rate of L929 cells in PA group was significantly increased （P<0.001） and the level of lipid deposition in the cells was significantly increased （P<0.001）. The TEM results showed that the mitochondria in L929 cells in PA group exhibited swelling， disruption of cristae structure， and fragmentation， while the number of MAMs contact sites was reduced and the coupling distance was increased. The flow cytometry， JC-1 method， and RT-qPCR method results showed that compared with control group， the level of ROS in L929 cells in PA group was significantly increased （P<0.001），the mitochondrial membrane potential of the cells was significantly decreased （P<0.01）， and the expression levels of MFN2 mRNA and protein in the cells were significantly decreased （P<0.01）. Conclusion PA-induced lipotoxicity downregulates MFN2 expression and disrupts MAMs structure， thereby promoting oxidative stress， mitochondrial depolarization， and cell apoptosis. These findings suggest that under PA-induced lipotoxic conditions， the downregulation of MFN2 expression and the damage to MAMs structure are closely associated with mitochondrial homeostasis imbalance and cell apoptosis.

Objective To investigate the ameliorative effect of Yijing Decoction （YJD） on ovarian reserve function in the naturally aged female Drosophila， and to elucidate its potential mechanism. Methods A total of 1 920 female Drosophila melanogaster were randomly divided into young group， older group， low dose of YJD （0.25 g·L^-1） group， medium dose of YJD （0.50 g·L^-1） group， high dose of YJD （1.00 g·L^-1） group， and positive drug 20-hydroxyecdysone （20E） group， with 320 Drosophila in each group. A hundred female Drosophila were selected from each group and male Drosophila were matched to each group at a ratio of 1∶2 （male∶female） and co-cultured for 7 d. Then the female Drosophila were collected and allowed to lay eggs alone for 48 h. The numbers of eggs laid and pupae formed were counted， and the pupation rates were calculated. Ovaries of the Drosophila in various groups were dissected under a stereomicroscope， and the lengths and diameters were measured to evaluate the morphology of the ovaries. Enzyme-linked immunosorbent assay （ELISA） method was used to detect the levels of 20E and juvenile hormone （JH） in ovarian tissues of the Drosophila in various groups. Immunofluorescence staining method was used to observe the numbers of germline stem cells （GSCs） in ovarian tissues of the Drosophila in various groups. Western blotting method was used to detect the expression levels of proteins related to the bone morphogenetic protein （BMP）/mothers against decapentaplegic homolog （Smad） signaling pathway， including Bam， Nos， Dpp， Smad4， Smad1/5/9， and phosphorylated Smad1/5/9 （p-Smad1/5/9）， in the ovarian tissue of the Drosophila in various groups. Results Compared with young group， the numbers of eggs laid and pupae formed as well as the pupation rate in older group were significantly decreased （P<0.01）， the ovarian morphology was atrophic and the ovarian length and diameter were significantly decreased （P<0.01）， the levels of 20E and JH in ovarian tissue of the Drosophila were significantly decreased （P<0.01）， and the number of GSCs in ovarian tissue of the Drosophila was significantly decreased （P<0.01）. Compared with older group， the numbers of eggs laid and pupae formed as well as the pupation rates in low dose of YJD group， medium dose of YJD group， high dose of YJD group， and 20E group were significantly increased （P<0.05 or P<0.01）， the ovarian recovered to full and normal morphology， the ovarian lengths and diameters of the Drosophila were significantly increased （P<0.05 or P<0.01）， the levels of 20E and JH in the ovarian tissue of the Drosophila were significantly increased （P<0.05 or P<0.01）； the numbers of GSCs in the ovarian tissue of the Drosophila in medium dose of YJD group， high dose of YJD group， and 20E group were significantly increased （P<0.01）. Compared with young group， the expression levels of Bam， Nos， Dpp， Smad4， and p-Smad1/5/9 proteins in the ovarian tissue of the Drosophila were significantly decreased in older group （P<0.01）. Compared with older group， the expression levels of Bam， Nos， Dpp， Smad4， and p-Smad1/5/9 proteins in the ovarian tissue of the Drosophila in low dose of YJD group， medium dose of YJD group， high dose of YJD group， and 20E groupwere significantly increased （P<0.05 or P<0.01）. Conclusion YJD ameliorates ovarian reserve function in older female Drosophila， enhances their reproductive capacity， restores ovarian morphology， up-regulates ovarian hormone levels， and increases the number of GSCs. Its mechanism is associated with regulating the BMP/Smad signaling pathway to promote GSC regeneration.

Objective To discuss the effect of co-expression of beta-2-microglobulin （B2M） protein and chimeric antigen receptor （CAR） molecule on the biological behaviors of mesothelin （MSLN）-targeted chimeric antigen receptor T cell（CAR-T）， and to clarify the role of B2M in enhancing the therapeutic efficacy of CAR-T cells against MSLN-positive solid tumors. Methods The B2M and CAR co-expressing third-generation anti-MSLN CAR-T （B2M-28bbz-T） cells and control traditional anti-MSLN CAR-T （28bbz-T） cells were constructed. The 293T cells stably overexpressing MSLN （293T-MSLN） were constructed. Real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of B2M， variable region of heavy chain （VH）， and variable region of light chain （VL） mRNA in the CAR-T cells in various groups. Flow cytometry was used to detect the expressions of B2M protein and CAR molecule in the cells and the proliferation abilities of the CAR-T cells in various groups. The 293T-MSLN cells and MSLN-positive ovarian cancer SK-OV-3 cells were selected as target cells. Target cells were co-incubated with untransduced T lymphocytes， 28bbz-T cells， and B2M-28bbz-T cells at an effector-to-target ratio （E/T） of 1∶1 for 72 h. After co-incubation， the target cells were divided into T group， 28bbz-T group， and B2M-28bbz-T group. Cytotoxicity assay was used to detect the activities of target cells in various groups under the action of CAR-T cells. Enzyme-linked immunosorbent assay （ELISA） method was used to detect the levels of interferon-γ （IFN-γ） and tumor necrosis factor-α （TNF-α） in the supernatants of target cells in various groups. Twenty female NSG mice were randomly divided into four groups： phosphate-buffered saline（PBS） group， 28bbz-T group， B2M-28bbz-T group， and blue fluorescent protein （BFP）-28bbz-T group， with 5 mice in each group. The SK-OV-3 cells were subcutaneously implanted into the mice to construct ovarian cancer cell line-derived xenograft （CDX） models. The corresponding CAR-T cells were injected into the mice in various groups via the tail vein on the 7th day. After the observation， the mice were sacrificed， and the tumor volumes and weights of the mice in various groups were measured. Results The B2M-28bbz-T cells and 28bbz-T cells were successfully constructed. The RT-qPCR method results showed that compared with 28bbz-T group， the expression level of B2M mRNA in B2M-28bbz-T group was significantly increased （P<0.01）， while no significant differences were observed in the expression levels of VH and VL mRNA （P>0.05）. The flow cytometry results showed that no significant differences were observed in CAR molecule expression and proliferation ability of cells between 28bbz-T group and B2M-28bbz-T group. The cell toxicity assay results showed that compared with 28bbz-T group， the activities of 293T-MSLN cells and SK-OV-3 cells in B2M-28bbz-T group were significantly decreased （P<0.01 or P<0.001）. The ELISA method results showed that compared with 28bbz-T group， the levels of IFN-γ and TNF-α in the supernatant of 293T-MSLN cells and SK-OV-3 cells in B2M-28bbz-T group were significantly increased （P<0.05）. The in vivo experiment results showed that compared with PBS group， the tumor growth was inhibited in the mice in 28bbz-T group and B2M-28bbz-T group， and the tumor of one mouse in B2M-28bbz-T group was completely cleared. Compared with 28bbz-T group， the tumor weight of the mice in B2M-28bbz-T group was significantly decreased （P<0.01）， while no statistically significant difference was observed in the tumor weight of the mice in BFP-28bbz-T group （P>0.05）. Conclusion Co-expression of B2M and CAR molecules may enhance the cytotoxicity and in vivo anti-tumor activity of MSLN-targeting CAR-T cells by promoting T lymphocyte activation and pro-inflammatory cytokine secretion.

Objective To investigate the inhibitory effect of the combination of Astragaloside Ⅳ（AST Ⅳ）， Panax notoginseng saponins （PNS）， and borneol （APB） on neuroinflammation after cerebral ischemia/reperfusion （CI/R） in the rats by modulating aerobic glycolysis and microglial polarization， and to elucidate its synergistic mechanism. Methods Twenty-five healthy male SD rats were selected and randomly divided into sham operation group，model group， low dose of APB group， high dose of APB group， and positive control drug butylphthalide （NBP） group， with 5 rats in each group. The rat models of focal cerebral ischemia were established by middle cerebral artery occlusion （MCAO） using the suture occlusion method， while those in sham operation group did not undergo vascular occlusion.The rats in various groups received intragastric administration twice daily， starting at the onset of reperfusion after 2 h of ischemia. The rats in low dose of APB group were given 60 mg·kg^-1 borneol+80 mg·kg^-1 AST Ⅳ+200 mg·kg^-1 PNS， the rats in high dose of APB group were given 120 mg·kg^-1 borneol+160 mg·kg^-1 AST Ⅳ+ 400 mg·kg^-1 PNS， and the rats in NBP group were given 80 mg·kg^-1 NBP. The rats in sham operation group and model group were given equal volumes of pure water. At 2 h before execution， the modified neurological severity score（mNSS） of the rats in various groups were assessed. At 48 h after reperfusion， the rats were anesthetized， and the blood was collected from the abdominal aorta， and then the rats were sacrificed to collect the ischemic side of brain tissue. HE staining and Nissl staining were used to observe the neuronal damage and the number of Nissl bodies in the hippocampal region of brain tissue in the rats in various groups. Enzyme-linked immunosorbent assay （ELISA） method was used to detect the serum levels of interleukin-1β （IL-1β）， interleukin-10 （IL-10）， tumor necrosis factor-α （TNF-α）， and interleukin-4（IL-4） in the rats in various groups. Immunofluorescence staining was used to detect the polarization state of microglia towards M1 and M2 phenotypes in the brain tissue of the rats in various groups. Western blotting method was used to detect the expression levels of the key glycolytic rate-limiting enzymes lactate dehydrogenase A （LDHA）， glucose transporter 1 （GLUT1）， and hexokinase 2 （HK2） proteins in the brain tissue of the rats in various groups. Results ?According to the mNSS score， compared with sham operation group， the neurological deficit score of the rats in model group was significantly increased （P<0.05）； compared with model group， the neurological deficit scores of the rats in low dose of APB group， high dose of APB group， and NBP group were significantly decreased （P<0.05 or P<0.01）. The HE staining results showed regularly arranged neurons in the hippocampal region of the rats in sham operation group， and pyknosis and vacuolar degeneration of neuronal nuclei were observed in the rats in model group. Compared with model group， the neuronal damage was alleviated in low-dose APB group， high-dose APB group， and NBP group， as indicated by a reduction in pyknotic and vacuolated cells. Compared with sham operation group， the neuronal survival rate in the hippocampal region of the rats in model group was significantly decreased （P<0.05）； compared with model group， the neuronal survival rates in the hippocampal regions of the rats in low dose of APB group， high dose of APB group， and NBP group were significantly increased （P<0.05）. The Nissl staining results showed that compared with sham operation group， the number of Nissl bodies in neurons in the hippocampal region of the rats in model group was significantly decreased （P<0.05）； compared with model group， the numbers of Nissl bodies in the hippocampal neurons of the rats in low dose of APB group， high dose of APB group， and NBP group were significantly increased （P<0.05）. The ELISA results showed that compared with sham operation group， the levels of pro-inflammatory factors IL-1β and TNF-α in serum in the rats in model group were significantly increased （P<0.05）， while the levels of IL-10 and IL-4 in serum were significantly decreased （P<0.05 or P<0.01）； compared with model group， levels of IL-1β and TNF-α in serum in the rats in low dose of APB group， high dose of APB group， and NBP group were significantly decreased （P<0.05）， while the levels of IL-10 and IL-4 in serum were significantly increased （P<0.05 or P<0.01）. The immunofluorescence staining results showed that compared with sham operation group， the co-expression fluorescence intensity of inducible nitric oxide synthase（iNOS）/ionized calcium-binding adapter molecule 1（Iba-1） in the brain tissue of the rats in model group was significantly increased （P<0.01）， while that of arginase-1（Arg-1）/Iba-1 was significantly decreased （P<0.01）； compared with model group， the co-expression fluorescence intensities of iNOS/Iba-1 in the brain tissue of the rats in low dose of APB group， high dose of APB group， and NBP group were significantly decreased （P<0.05 or P<0.01）， while those of Arg-1/Iba-1 were significantly increased （P<0.05 or P<0.01）. The Western blotting results showed that compared with sham operation group， the expression levels of LDHA， GLUT1， and HK2 proteins in the brain tissue of the rats in model group were significantly increased （P<0.05）； compared with model group， the expression levels of LDHA protein in the brain tissue of the rats in low dose of APB group and high dose of APB group were significantly decreased （P<0.05）， while the expression levels of GLUT1 and HK2 proteins in high dose of APB group and NBP group were significantly decreased（P<0.05）. Conclusion ?APB can inhibit the polarization of microglia towards the M1 phenotype by downregulating the expression of glycolytic enzymes LDHA， GLUT1， and HK2 proteins， thereby alleviating inflammatory responses in brain tissue and improving neurological function in the rats after CI/R.

Objective To investigate the effect of pyraclostrobin on mitochondrial function and dynamic balance in mouse spermatocyte GC-2 cells， and to elucidate its possible mechanism. Methods Methylthiazolyldiphenyl-tetrazolium bromide（MTT） method was used to detect the cell activity of GC-2 cells in various groups after treatment with different concentrations of pyraclostrobin， so as to determine the appropriate treatment concentration. Mouse spermatocyte cell line GC-2 was divided into control group， low dose of pyraclostrobin （0.8 μmol·L^-1） group， medium dose of pyraclostrobin （1.6 μmol·L^-1） group， and high dose of pyraclostrobin （2.4 μmol·L^-1） group. JC-1 mitochondrial membrane potential assay kit was used to detect the mitochondrial transmembrane potential difference （ΔΨ_m）across the inner membrane of mitochondrial in GC-2 cells in various groups. 2'，7'-dichlorodihydrofluorescein diacetate （DCFH-DA） fluorescent probe method was used to detect the levels of reactive oxygen species （ROS） in GC-2 cells in various groups. Adenosine triphosphate （ATP） assay kit was used to measure the levels of ATP in GC-2 cells in various groups. Mito-tracker fluorescence probe method was used to detect the fusion and fission of mitochondria in GC-2 cells in various groups. Western blotting method was used to detect the expression levels of mitochondrial fusion and fission related proteins， optic atrophy 1 （OPA1） and dynamin-related protein 1 （DRP1）， in GC-2 cells in various groups. Results Compared with control group， the cell activities of GC-2 cells were significantly decreased in groups with pyraclostrobin concentration≥1 μmol·L^-1 （P<0.001）. The half maximal inhibitory concentration （IC₅₀） of pyraclostrobin against GC-2 cells was 4.8 μmol·L^-1. Compared with the control group， the ΔΨ_m in the GC-2 cells were significantly decreased in low dose of pyraclostrobin group， medium dose of pyraclostrobin group， and high dose of pyraclostrobin group （P<0.05 or P<0.01），the levels of ROS in the GC-2 cells were significantly increased（P<0.05 or P<0.01），and the ATP levels in the GC-2 cells were significantly decreased （P<0.001）. The Mito-Tracker staining results showed that the mitochondria in GC-2 cells in control group were distributed in a continuous network pattern with good fusion， while those in low dose of pyraclostrobin group， medium dose of pyraclostrobin group， and high dose of pyraclostrobin group showed fragmented morphology with obvious network structure breakage， decreased fusion， and increased fission. The Western blotting results showed that compared with control group， the expression levels of OPA1 protein in the GC-2 cells were decreased in medium dose of pyraclostrobin group and high dose of pyraclostrobin group （P<0.05 or P<0.01）， and the expression levels of DRP1 protein in the GC-2 cells were significantly increased in low dose of pyraclostrobin group，medium dose of pyraclostrobin group， and high dose of pyraclostrobin group （P<0.001）. Conclusion Pyraclostrobin induced an imbalance between mitochondrial fusion and fission in GC-2 cells， leading to mitochondrial dysfunction. Its toxic effect may be associated with abnormal regulation of mitochondrial dynamics.

Objective To investigate the effect of the NIPBL gene knockdown on osteogenic differentiation of the mouse bone marrow mesenchymal stem cells （BMSCs）， and to elucidate the underlying mechanism through the sonic hedgehog （Shh） signaling pathway. Methods The BMSCs isolated from C57 mice were cultured in vitro to the third generation. Flow cytometry was used to identify the expressions of cell surface markers on the BMSCs. Lentivirus carrying green fluorescence protein （GFP） was used to construct the BMSC model with stable NIPBL knockdown. The cells were divided into control group（routine culture）， sh-NC group （transfected with Scramble shRNA lentivirus）， and sh-NIPBL group（transfected with NIPBL shRNA lentivirus）. Real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of NIPBL mRNA in the BMSCs in various groups. The BMSCs in various groups were subjected to osteogenic induction， and the morphological changes were dynamically observed. The cells were collected on days 7， 14， and 21 after induction. Western blotting method was used to detect the expression levels of Shh signaling pathway proteins， including Shh， patched homolog 1 （PTCH1）， and glioma-associated oncogene homolog 1 （GLI1）， as well as osteogenic markers， including alkaline phosphatase （ALP）， osteocalcin （OCN）， and collagen type Ⅰ alpha 1 chain （COL1A1） proteins in the cells in various groups. Alizarin red staining was used to assess the mineralization capacities of the BMSCs in various groups on day 21 after induction. Results Third-generation BMSCs exhibited good growth status， showing a long-spindle or fusiform morphology and the typical CD105?CD45? phenotype. The stable GFP expression rates in the BMSCs of the mice in sh-NIPBL group and sh-NC group exceeded 90%， while no fluorescence signal was observed in control group. Compared with control group， the expression level of NIPBL mRNA in the BMSCs in sh-NIPBL group was significantly decreased （P<0.001）， indicating successful establishment of the NIPBL-knockdown model. On day 21 after osteogenic induction， dense mineralized nodules were observed in control group， while only a small number of scattered， tiny， sparse， and non-fused mineralized nodules were seen in sh-NIPBL group， suggesting delayed osteogenic differentiation. On day 21 after induction， compared with sh-NC group， the expression levels of Shh， PTCH1， GLI1， COL1A1， ALP， and OCN proteins in the BMSCs in sh-NIPBL group were significantly decreased （P<0.05）. On days 7， 14， and 21 after induction， the expression levels of aforementioned proteins in the BMSCs in sh-NIPBL group were consistently lower than those in control group and sh-NC group. The Alizarin red staining results showed a large number of dense， fused orange-red mineralized nodules in the BMSCs in control group and sh-NC group， while only scattered orange-red spots in the BMSCs were observed in sh-NIPBL group. Compared with control group， the mineralization capacity of cells in sh-NIPBL group was significantly decreased （P<0.01）. Conclusion Knockdown of the NIPBL gene may reduce the osteogenic differentiation capacity and mineralization capacity of mouse BMSCs， and its mechanism is related to inhibiting the activity of the Shh signaling pathway and downregulating the expression of osteogenesis-related proteins.

Objective To discuss the effect of thioredoxin domain-containing protein 9 （TXNDC9） gene knockdown on the apoptosis and autophagy of colorectal cancer （CRC） cells， and to clarify its potential molecular mechanism based on the p53-dependent pathway. Methods The human normal colonic epithelial cell line NCM460 and colon cancer cell lines HCT116， SW620， SW480， and LoVo were selected. The HCT116 cells were divided into NC group （transfected with negative control si-NC）， si-TXNDC9 group （transfected with si-TXNDC9）， and p53 pathway inhibitor PTF-α group （PTF-α group， transfected with si-TXNDC9 and treated with 5 μmol·L^-1 PTF-α for 24 h）. Plate colony formation assay and Transwell chamber assay were used to detect the numbers of clone formation and invasion cells in various groups. Western blotting method was used to detect the expression levels of autophagy， apoptosis， and p53 pathway-related proteins in the cells in various groups. Terminal deoxynucleotidyl transferase dUTP nick end labeling （TUNEL） assay was used to detect the apoptotic rates of the cells in various groups.Transmission electron microscope （TEM） was used to observe the numbers of autophagosomes in the cells in various groups. The xenograft tumor models in nude mice were established， and the mice were randomly divided into control group （inoculated with si-NC-transfected HCT116 cells）， knockdown group （inoculated with si-TXNDC9-transfected HCT116 cells）， and inhibitor group （inoculated with si-TXNDC9-transfected HCT116 cells and intraperitoneally injected with PTF-α） （n=5）. The tumor volumes of nude mice in various groups were measured and recorded every 7 days， and the mice were sacrificed on the 28th day， and the tumors were stripped and weighed. Results In cell experiments， compared with the NCM460 cells， the expression levels of TXNDC9 protein in the HCT116， SW620， SW480， and LoVo cells were significantly increased （P<0.05）. Compared with NC group， the numbers of clone formation and invasion cells of HCT116 cells in si-TXNDC9 group were significantly decreased （P<0.05）， the apoptotic rate of the cells was significantly increased （P<0.05）， the expression levels of B-cell lymphoma 2（Bcl-2） and B-cell lymphoma-extra large （Bcl-XL） proteins in the cells were significantly decreased （P<0.05）， the expression level of Bcl-2-associated X protein （BAX） was significantly increased （P<0.05）， the number of autophagosomes was significantly increased （P<0.05）， the expression levels of autophagy-related protein 7 （ATG7） and Beclin-1 proteins and the ratio of microtubule-associated protein 1 light chain 3 （LC3）-Ⅱ/LC3-Ⅰ were significantly increased （P<0.05）， the expression level of P62 protein was significantly decreased （P<0.05）， and the expression levels of P53 and P21 proteins were significantly increased （P<0.05）. Compared with si-TXNDC9 group， the numbers of clone formation and invasion cells of HCT116 cells in PTF-α group were significantly increased （P<0.05）， the apoptotic rate of the cells was significantly decreased （P<0.05）， the expression levels of Bcl-2 and Bcl-XL proteins in the cells were significantly increased （P<0.05）， the expression level of BAX protein was significantly decreased （P<0.05）， the number of autophagosomes was significantly decreased （P<0.05）， the expression levels of ATG7 and Beclin-1 proteins and the LC3-Ⅱ/LC3-Ⅰ ratio in the cells were significantly decreased （P<0.05）， the expression level of P62 protein was significantly increased （P<0.05）， and the expression levels of P53 and P21 proteins were significantly decreased （P<0.05）. In animal experiments， compared with control group， the tumor weights and tumor volumes at each time point in knockdown group were significantly decreased （P<0.05）； compared with knockdown group， the tumor weights and tumor volumes at each time point in inhibitor group were significantly increased （P<0.05）. Conclusion TXNDC9 knockdown can promote apoptosis and autophagy in the CRC cells by regulating the p53-dependent pathway， thereby inhibiting their proliferation and invasion abilities.

Objective To discuss the effects of Panax notoginseng saponins （PNS） on the biological behavior of gastric cancer （GC） cells and to analyze its relationship with the Kelch-like ECH-associated protein 1 （Keap1）-nuclear factor E2-related factor 2 （Nrf2） signaling pathway. Methods GC cell lines AGS and HGC27 cells and human normal gastric mucosal epithelial cells GES-1 were used as the research objects and treated with different concentrations of PNS. MTT method was used to detect the cell activity in various groups. AGS and HGC27 cells were divided into control group （0 μmol·L^-1 PNS group） and 5， 10， and 20 μmol·L^-1 PNS groups. Colony formation assay was used to detect the proliferation abilities of the cells in various groups. Transwell chamber assay was used to detect the numbers of migration cells and invasion cells in various groups. 2'，7'-dichlorodihydrofluorescein diacetate （DCFH-DA） fluorescent probe method was used to detect the reactive oxygen species （ROS） levels in the cells in various groups. Western blotting method was used to detect the expression levels of apoptosis-related proteins B-cell lymphoma 2 （Bcl-2）， Bcl-2-associated X protein （Bax）， Bcl-2-associated death promoter （Bad）， cysteinyl aspartate-specific proteinase 3 （Caspase-3）， and cleaved Caspase-3， as well as Keap1 and Nrf2 proteins in the cells in various groups. Co-immunoprecipitation （Co-IP） method combined with Western blotting method was used to detect the interaction between Keap1 and Nrf2 and Nrf2 ubiquitination in the cells in various groups. Results After treatment for 48 h， compared with control group， the activities of AGS and HGC27 cells in 5， 10， and 20 μmol·L^-1 PNS groups were significantly decreased （P<0.001）， while there was no statistically significant difference in the activity of GES-1 cells （P>0.05）. Compared with control group， the colony formation numbers， numbers of migration cells， and numbers of invasion cells of AGS and HGC27 cells in 5， 10， and 20 μmol·L^-1 PNS groups were significantly decreased （P<0.001）， while the apoptotic rates and the ROS levels in the cells in 5， 10， and 20 μmol·L^-1 PNS groups were significantly increased （P<0.05 or P<0.01）. Compared with control group， the expression levels of Bcl-2 and Nrf2 proteins in AGS and HGC27 cells in 5， 10， and 20 μmol·L^-1 PNS groups were significantly decreased （P<0.001）， while the expression levels of Bad， Bax， cleaved Caspase-3， and Keap1 proteins were significantly increased （P<0.001）. Compared with control group， the co-precipitation signals of Keap1 and Nrf2 in AGS and HGC27 cells in PNS group were increased， while the ubiquitination signals of Nrf2 in AGS and HGC27 cells were decreased. Conclusion PNS inhibits the proliferation， migration， and invasion of GC cells， promotes cell apoptosis， and enhances intracellular oxidative stress level. Its effect is related to the regulation of the expression of Keap1-Nrf2 signaling pathway-related proteins and the interaction between Keap1 and Nrf2.

Objective To systematically evaluate the developmental toxicity and potential cardiac and hepatic toxicity of the aqueous extract of Asarum heterotropoides based on zebrafish embryo model， to clarify its dose-response relationship and sensitive target organs， and to provide experimental evidence for the safety assessment of Asarum heterotropoides. Methods Wild-type AB strain zebrafish embryos at 1 day post-fertilization （dpf） were selected. After excluding unfertilized and abnormally developed individuals， they were randomly divided into a control group （culture medium only） and groups treated with different concentrations of the aqueous extract of Asarum heterotropoides （100， 200， 300， 500， 1 000， and 2 000 mg·L?1）， with 18 embryos in each group. Exposure lasted until 5 dpf. The hatching rate at 2 dpf and mortality rate at 5 dpf of zebrafish embryos in various groups were recorded. Concentration-effect curves were plotted， and Probit regression analysis was performed to calculate the median lethal concentration （LC₅₀）， as well as 20% lethal concentration （LC₂₀）， 10% lethal concentration（LC₁₀）， and 5% lethal concentration（LC₅）. At 5 dpf， morphological changes in zebrafish embryos in various groups were observed. Heart rate， sinus venosus-bulbus arteriosus （SV-BA） distance， and pericardial area of zebrafish embryos in various groups were measured to evaluate embryonic cardiac structure and function. Oil Red O staining was used to determine the hepatic integrated optical density （IOD） values to assess the levels of lipid accumulation in the liver of zebrafish embryos in various groups. Results Compared with control group， the hatching rates of zebrafish embryos in 300 mg·L^-1 group and higher concentration groups were significantly decreased （P<0.05）， while the mortality rates were significantly increased （P<0.01）， in a concentration-dependent manner. Probit regression analysis results showed that the LC₅₀ of the aqueous extract of Asarum heterotropoides was 641.11 mg·L^-1， and the LC₂₀， LC₁₀， and LC₅ were 317.78， 223.33， and 146.67 mg·L^-1， respectively. Morphological observation revealed that the zebrafish embryos in 200 mg·L^-1 group and higher concentration groups exhibited abnormalities such as delayed yolk sac absorption， pericardial edema， jaw malformation， and shortened body length. Compared with control group， the heart rates of zebrafish embryos in LC₁₀ group and LC₂₀ group were significantly decreased （P<0.01）， while the SV-BA distances and pericardial areas of zebrafish embryos were significantly increased （P<0.01）， indicating impaired cardiac structural development and functional status. The Oil Red O staining results showed increased lipid deposition in the liver of the zebrafish embryos in LC₅ group， LC₁₀ group and LC₂₀ group， and the levels of lipid accumulation in the liver were significantly higher than that in control group （P<0.01）， among which LC₂₀ group showed the highest degree of lipid deposition in the liver. Conclusion The aqueous extract of Asarum heterotropoides showed concentration-dependent developmental toxicity on zebrafish embryos and may induce cardiac structural and functional abnormalities as well as increased hepatic neutral lipid deposition at doses below the LC₅₀. The results suggested that the heart and liver may be its sensitive target organs.

Objective To investigate the inhibitory effect of ginseng-lithospermum erythrorhizon extract on the proliferation and invasion of human keloid fibroblasts （HKF） based on network pharmacology， molecular docking technology and in vitro cell experiments， and to clarify its mechanism. Methods Traditional Chinese Medicine Systems Pharmacology （TCMSP） database， Traditional Chinese Medicine Integrated Database （TCMID）， Swiss Target Prediction database， and UniProt database were used to retrieve the active components of ginseng-lithospermum extract and their corresponding target genes. GeneCards database and Online Mendelian Inheritance in Man （OMIM） database were used to screen keloid （KD）-related targets. STRING online platform and Cytoscape 3.10.3 software were used to construct the protein-protein interaction （PPI） network and identify core targets. Metascape database was used to perform Gene Ontology （GO） functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathway enrichment analysis on the above core targets. CB-Dock2 website was used to conduct molecular docking analysis and visualize the results. HKF cells were divided into blank group （100 μL complete medium）， control group （100 μL complete medium supplemented with equal volume of solvent）， and low dose， medium dose， and high dose of ginseng-lithospermum erythrorhizon extract groups （100 μL complete medium supplemented with ginseng-lithospermum extract at final concentrations of 10， 20， and 40 mg·L^-1， respectively）. Cell counting kit-8 （CCK-8） method was used to detect the survival rates of HKF cells in various groups. Flow cytometry method was used to detect the percentages of cells in different cell cycle phases in various groups. Transwell chamber assay was used to detect the numbers of invasion cells in HKF cells in various groups. Real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of serine/threonine protein kinase 1 （AKT1）， interleukin-1β（IL-1β）， and prostaglandin-endoperoxide synthase 2 （PTGS2） mRNA in HKF cells in various groups. Results A total of 27 active components of ginseng-lithospermum extract and 31 intersection targets between the drug and KD were screened out， among which 9 core targets were identified， including IL-1β， AKT1 and PTGS2. GO functional enrichment analysis mainly involved biological processes including cell proliferation， apoptosis， and oxidative stress response. KEGG signaling pathway enrichment analysis mainly involved phosphatidylinositol 3-kinase/protein kinase B （PI3K-Akt） signaling pathway， mitogen-activated protein kinase （MAPK） signaling pathway， Janus kinase/signal transducer and activator of transcription （JAK/STAT） signaling pathway， and cancer-related pathways. The binding energies between PTGS2 and the five active components were all lower than -7 kcal·mol^-1. Compared with control group， the survival rates of HKF cells in low dose， medium dose， and high dose of ginseng-lithospermum erythrorhizon extract groups were significantly decreased （P<0.05）， the percentages of cells in G₀/G₁ phase were significantly increased （P<0.05）， and the numbers of invasion cells were significantly decreased （P<0.05）. Compared with control group， the expression levels of AKT1 and IL-1β mRNA in HKF cells in low dose， medium dose， and high dose of ginseng-lithospermum erythrorhizon extract groups were significantly decreased （P<0.05）， while the expression levels of PTGS2 mRNA were significantly increased （P<0.05 or P<0.01）. Conclusion Ginseng-lithospermum erythrorhizon extract inhibited the proliferation and invasion of HKF cells and induced cell cycle arrest. Its mechanism may be associated with regulating the core targets AKT1， IL-1β， and PTGS2， and mediating PI3K-Akt， MAPK， and JAK/STAT signaling pathways.

Objective To investigate the improvement effect of ginsenoside Rg3 （G-Rg3） on high glucose-induced injury in human renal tubular epithelial HK2 cells， and to elucidate its mechanism. Methods A high glucose-induced injury model of human renal tubular epithelial HK2 cells was established using high glucose（HG） medium （containing 25 mmol·L^-1 glucose）. Cells were divided into negative control （NC） group （medium containing 5.5 mmol·L^-1 glucose）， HG group （HG medium）， nuclear factor erythroid 2-related factor 2 （Nrf2） inhibitor ML385 group （ML385 group） （HG medium+5 μmol·L^-1 ML385）， G-Rg3 intervention group （Rg3 group） （HG medium+40 μmol·L^-1 G-Rg3）， and ML385+Rg3 group （HG medium + 5 μmol·L^-1 ML385 + 40 μmol·L^-1 G-Rg3）. Cell counting kit-8 （CCK-8） method was used to detect the cell activities in cells in various groups. 2'，7'-dichlorodihydrofluorescein diacetate（DCFH-DA） fluorescence method was used to detect the levels of reactive oxygen species （ROS） in the cells in various groups. Commercial kits were used to detect the activities of superoxide dismutase （SOD） and lactate dehydrogenase （LDH） and the levels of malondialdehyde （MDA） and ferrous ion（Fe²⁺） in the cells in various groups. Western blotting method was used to detect the expression levels of Nrf2， heme oxygenase-1 （HO-1）， glutathione peroxidase 4 （GPX4）， and acyl-CoA synthetase long-chain family member 4 （ACSL4） proteins in the cells in various groups. Results The results of CCK-8 method， DCFH-DA fluorescence method and kit method showed that， after 48 h of culture， compared with NC group， the cell activity was significantly decreased in HG group （P<0.01），the level of ROS， the activity of LDH， and the levels of MDA and Fe²⁺ in the cells were significantly increased （P<0.01）， and the activity of SOD was significantly decreased （P<0.01）. Compared with HG group， the level of ROS， the activity of LDH， and the levels of MDA and Fe²⁺ in the cells in ML385 group were significantly increased （P<0.05）， and the activity of SOD was significantly decreased （P<0.05）； the cell activity in Rg3 group was significantly increased （P<0.01）， the level of ROS， the activity of LDH， and the levels of MDA and Fe²⁺ in the cells were significantly decreased （P<0.01）， and the activity of SOD was significantly increased （P<0.01）. Compared with Rg3 group， the cell activity in ML385+Rg3 group was significantly decreased （P<0.01）， the level of ROS， the activity of LDH， and the levels of MDA and Fe²⁺ in the cells were significantly increased （P<0.01）， and the activity of SOD was significantly decreased （P<0.01）. The results of Western blotting method showed that， compared with NC group， the expression levels of Nrf2， HO-1， and GPX4 proteins in HK2 cells were significantly decreased in HG group （P<0.01）， and the expression level of ACSL4 protein was significantly increased （P<0.01）. Compared with HG group， the expression level of GPX4 protein in the cells in ML385 group was significantly decreased （P<0.01）， and the expression level of ACSL4 protein was significantly increased （P<0.01）； the expression levels of Nrf2， HO-1， and GPX4 proteins in HK2 cells were significantly increased in Rg3 group （P<0.01）， and the expression level of ACSL4 protein was significantly decreased （P<0.05）. Compared with Rg3 group， the expression levels of Nrf2， HO-1， and GPX4 proteins in HK2 cells in ML385+Rg3 group were significantly decreased （P<0.01）， and the expression level of ACSL4 protein was significantly increased （P<0.05）. Conclusion ?G-Rg3 showed an ameliorative effect on HG-induced oxidative injury in human renal tubular epithelial cells， and its mechanism may be associated with upregulating Nrf2 expression， activating the Nrf2/HO-1/GPX4 signaling pathway， and inhibiting ferroptosis.

Objective To discuss the therapeutic effect of Mishitong capsule on calculi-inducing solution-induced kidney stone mice，and to clarify its mechanism. Methods A total of 50 SPF-grade male ICR mice were selected. Forty mice were randomly selected and intragastrically administered with 3 mL·d^-1 calculi-inducing solution （0.75% ethylene glycol+0.75% ammonium chloride） for 2 weeks to establish the kidney stone mouse model， and the remaining 10 mice served as control group. After successful modeling， the 40 model mice were randomly divided into model group， low dose of Mishitong capsule group， high dose of Mishitong capsule group， and positive drug group， with 10 mice in each group. One week after modeling， the mice in low dose of Mishitong capsule group and high dose of Mishitong capsule group were intragastrically administered with 225 and 450 mg·kg^-1 Mishitong capsule suspension， respectively； the mice in positive drug group were intragastrically administered with 5 g·kg^-1 Paishi granule suspension； the mice in control group and model group were intragastrically administered with an equal volume of normal saline. The administration was performed for 2 consecutive weeks， during which the body mass of the mice was measured weekly. After administration， the mice in various groups were sacrificed， renal tissue was isolated， and serum and feces were collected. Kit method was used to detect the levels of reduced glutathione （GSH） and malondialdehyde （MDA） and the activity of superoxide dismutase （SOD） in renal tissue of the mice in various groups， as well as the levels of blood urea nitrogen （BUN）， creatinine （Cr）， calcium （Ca）， phosphorus （P）， and magnesium （Mg） in serum of the mice in various groups. Western blotting method was used to detect the expression levels of nuclear factor erythroid 2-related factor 2 （Nrf2）， autophagy-related protein 5 （ATG5）， P62， and heme oxygenase-1 （HO-1） proteins in renal tissue of the mice in various groups； 16S ribosomal RNA （rRNA） sequencing method was used to analyze the gut microbiota diversity of the mice in various groups. Results Compared with control group， the mice in model group showed a slight decrease in body mass in the early modeling stage， accompanied by mental lethargy and piloerection. Compared with model group， the declining trend of body mass was slowed， and the mental status was improved in the mice in low dose of Mishitong capsule group and high dose of Mishitong capsule group. The kit method results showed that compared with model group， the GSH levels and SOD activities in the renal tissue of the mice in low dose of Mishitong capsule group， high dose of Mishitong capsule group， and positive drug group were significantly increased （P<0.05）， and the MDA levels were significantly decreased （P<0.05）. Compared with model group， the BUN and Ca levels in the serum of the mice in low dose of Mishitong capsule group and high dose of Mishitong capsule group were significantly decreased （P<0.05 or P<0.01）， and the Mg level was significantly increased （P<0.05）； the Cr and P levels in the serum of the mice in high dose of Mishitong capsule group were significantly increased （P<0.05）； the BUN， Cr， Ca， and P levels in the serum of the mice in positive drug group were significantly decreased （P<0.05 or P<0.01）， and the Mg level was significantly increased （P<0.05）. The Western blotting results showed that compared with model group， the expression levels of HO-1， Nrf2， and ATG5 proteins in the renal tissue of the mice in low dose of Mishitong capsule group， high dose of Mishitong capsule group， and positive drug group were significantly increased （P<0.05）， and the expression levels of P62 protein were significantly decreased （P<0.05）. The 16S rRNA sequencing results showed that compared with model group， the Chao1 index and Pielou’s evenness index of gut microbiota of the mice in high dose of Mishitong capsule group and positive drug group were significantly increased （P<0.05）. At the phylum level， compared with model group， the relative abundance of Bacteroidetes in gut microbiota of the mice in low dose of Mishitong capsule group and high dose of Mishitong capsule group was decreased， and the relative abundances of Firmicutes and Proteobacteria were increased； at the genus level， compared with model group， the relative abundance of Prevotella in gut microbiota of the mice in high dose of Mishitong capsule group was increased， the relative abundance of Bacteroides was decreased， and a new dominant genus Helicobacter emerged. Conclusion Mishitong capsule ameliorates the pathophysiological damage in calculi-inducing solution-induced kidney stone mice， and its mechanism is potentially associated with reducing serum BUN and Cr levels， modulating serum Ca， P， and Mg levels， alleviating oxidative stress injury， regulating the expression of proteins related to the Nrf2/ATG5/P62 signaling pathway， and improving the gut microbiota structure.

Objective To investigate the improvement effect of Panax notoginseng saponins （PNS） on neurological injury in the rats with ischemic stroke by regulating aerobic glycolysis， and to elucidate its mechanism. Methods Forty-eight rats were randomly divided into sham operation group， model group， PNS group （40 mg·kg^-1）， and PNS+phosphatidylinositol 3-kinase （PI3K） inhibitor LY294002 group （5 mg·kg^-1） PNS+LY294002 group， with 12 rats in each group. Except for sham operation group， the rats in other groups were subjected to middle cerebral artery occlusion/reperfusion （MCAO/R） method to establish ischemic stroke models. Longa scoring method was used to evaluate the neurological deficit of the rats in various groups. Triphenyltetrazolium chloride （TTC） staining method was used to detect the percentages of cerebral infarction area of the rats in various groups. HE staining and Nissl staining methods were used to observe the pathological changes and neuronal morphology in the brain tissue of the rats in various groups. Enzyme-linked immunosorbent assay （ELISA） method was used to detect the levels of lactate dehydrogenase A （LDHA）， interleukin （IL）-1β， and IL-18 in the brain tissue of the rats in various groups. Real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of hexokinase （HK）1， HK2， phosphofructokinase（PFK）1， PFK2， and pyruvate kinase M2 （PKM2） mRNA in the brain tissue of the rats in various groups. Immunofluorescence staining method was used to detect the levels of reactive oxygen species （ROS） in the cells and the perimeter/area （P/A） ratio of ionized calcium-binding adapter molecule 1 （Iba1）-positive microglia in the brain tissue of the rats in various groups. Terminal deoxynucleotidyl transferase dUTP nick end labeling（TUNEL） method was used to detect the neuronal apoptosis rates in the brain tissue of the rats in various groups. Immunohistochemistry method and Western blotting method were used to detect the expression levels of PI3K/protein kinase B （Akt） pathway-related proteins and the ratios of phosphorylated PI3K （p-PI3K）/PI3K and phosphorylated Akt （p-Akt）/Akt in the brain tissue of the rats in various groups. Results Compared with sham operation group， the neurological deficit score of the rats in model group was significantly increased （P<0.05）， the percentage of cerebral infarction area was significantly increased （P<0.05）， and severe pathological damage was observed in the brain tissue of the rats； compared with model group， the neurological deficit scores of the rats in PNS group and PNS+LY294002 group were significantly decreased （P<0.05）， the percentages of cerebral infarction area were significantly decreased（P<0.05）， and the pathological damages in the brain tissue of the rats were alleviated； compared with PNS group， the neurological deficit score of the rats in PNS+LY294002 group was significantly increased （P<0.05）， the percentage of cerebral infarction area was significantly increased （P<0.05）， and the pathological damage in brain tissue was aggravated. The ELISA method results showed that compared with sham operation group， the levels of LDHA， IL-1β， and IL-18 in the brain tissue of the rats in model group were significantly increased （P<0.05）. Compared with model group， the levels of LDHA， IL-1β， and IL-18 in the brain tissue of the rats in PNS group and PNS+LY294002 group were significantly decreased （P<0.05）； compared with PNS group， the levels of LDHA， IL-1β， and IL-18 in the brain tissue of the rats in PNS+LY294002 group were significantly increased （P<0.05）. The results of RT-qPCR method showed that compared with sham operation group， the expression levels of HK1， HK2， PFK1， PFK2， and PKM2 mRNA in the brain tissue of the rats in model group were significantly decreased （P<0.05）. Compared with model group， the expression levels of the HK1， HK2， PFK1， PFK2， and PKM2 mRNA in the brain tissue of the rats in PNS group and PNS+LY294002 group were significantly increased （P<0.05）； compared with PNS group， the expression levels of the HK1， HK2， PFK1， PFK2， and PKM2 mRNA in the brain tissue of the rats in PNS+LY294002 group were significantly decreased （P<0.05）. The immunofluorescence staining results showed that compared with sham operation group， the level of ROS in cells in the brain tissue of the rats in model group was significantly increased （P<0.05）， and the P/A ratio of Iba1-positive microglia in the brain tissue of the rats was significantly decreased（P<0.05）； compared with model group， the levels of ROS in the cells in the brain tissue of the rats in PNS group and PNS+LY294002 group were significantly decreased （P<0.05）， and the P/A ratios of Iba1-positive microglia in the brain tissue of the rats were significantly increased （P<0.05）； compared with PNS group， the level of ROS in the cells in the brain tissue of the rats in PNS+LY294002 group was significantly increased （P<0.05）， and the P/A ratio of Iba1-positive microglia in the brain tissue of the rats was significantly decreased （P<0.05）. The TUNEL method results showed that compared with sham operation group， the neuronal apoptosis rate in the brain tissue of the rats in model group was significantly increased （P<0.05）； Compared with model group， the neuronal apoptosis rates in the brain tissue of the rats in PNS group and PNS+LY294002 group were significantly decreased （P<0.05）； compared with PNS group， the neuronal apoptosis rate in the brain tissue of the rats in PNS+LY294002 group was significantly increased （P<0.05）. The Western blotting method results showed that compared with sham operation group， the ratios of p-PI3K/PI3K and p-Akt/Akt in the brain tissue of the rats in model group were significantly decreased （P<0.05）； compared with model group， the ratios of p-PI3K/PI3K and p-Akt/Akt in the brain tissue of the rats in PNS group and PNS+LY294002 group were significantly increased （P<0.05）； compared with PNS group， the ratios of p-PI3K/PI3K and p-Akt/Akt in the brain tissue of the rats in PNS+LY294002 group were significantly decreased （P<0.05）. Conclusion PNS can effectively improve neurological deficit in the MCAO/R rats and reduce the level of ROS in brain tissue of the rats. Its mechanism may be related to activating the PI3K/Akt pathway， enhancing glycolytic capacity， and thereby promoting metabolic reprogramming of microglia.

Objective To investigate the protective effect of active components of Jingkang Capsule on cerebral ischemia injury in rats，and to elucidate its possible mechanism. Methods Seventy SD rats were randomly divided into sham operation group， model group， and low dose （252 mg·kg?1）， medium dose （630 mg·kg?1）， and high dose of Jingkang Capsule （1 260 mg·kg?1） groups. The drugs were prepared with normal saline and administered to the rats via daily oral gavage for 14 consecutive days as preventive treatment. The rats in sham operation group and model group were given equal volumes of normal saline. After 14 days of treatment， the rats in sham operation group underwent only vessel separation without any further treatment， while the rats in other groups were subjected to cerebral ischemia model establishment using the suture occlusion method. Longa scoring method was used to assess neurological deficit in the rats in various groups. Triphenyltetrazolium chloride （TTC） staining was used to detect the cerebral infarction area in the rats in various groups. Hematoxylin-eosin （HE） staining and Nissl staining were used to observe pathomorphological changes in the brain tissues of rats in various groups. A “drug-component-target” network was constructed， and bioinformatics was used to screen the action targets of Jingkang Capsule. Western blotting method was used to detect the expression levels of protein kinase B （Akt） and apoptosis-related proteins in the cortical tissue of the rats in various groups. Results The Longa scoring method results showed that compared with sham operation group， the rats in model group showed significant neurological deficits， and the neurological deficit score was significantly increased （P<0.001）. Compared with model group， the neurological deficit scores in medium and high doses of Jingkang Capsule groups were significantly decreased （P<0.05 or P<0.01）. The TTC staining results showed that compared with sham operation group， obvious cerebral infarction areas were observed in the brain tissue of the rats in model group， and the percentage of cerebral infarction area was significantly increased （P<0.001）. Compared with model group， the percentages of cerebral infarction area of the rats in medium dose and high dose of Jingkang Capsule groups were significantly decreased （P<0.05 or P<0.01）. HE staining and Nissl staining results showed that compared with sham operation group， the brain tissue on the ischemic side of the rats in model group had looser structure with visible neuronal pyknosis， and the number of Nissl bodies in neurons was significantly decreased （P<0.001）. Compared with model group， the brain tissue on the ischemic side of rats in medium dose of Jingkang Capsule group showed reduced neuronal deformation， orderly arrangement， and relatively intact structure， and the number of Nissl bodies in neurons was significantly increased （P<0.01）. A “drug-component-target” network was constructed using drug-component-intersection target data， and the top 10 targets were screened as core targets using the cytoHubba plugin. The Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analyses results showed that most enriched pathways of drug-disease intersection targets were related to apoptosis and inflammation signaling pathways. The Western blotting method results showed that compared with sham operation group， the ratio of phosphorylated Akt （p-Akt）/Akt in the brain tissue of the rats in model group was significantly decreased （P<0.01）， the expression level of B-cell lymphoma 2 （Bcl-2） protein was significantly decreased （P<0.01）， and the expression levels of Bcl-2-associated X protein （Bax） and cleaved cysteinyl aspartate-specific proteinase 3 （Cleaved Caspase-3） proteins were significantly increased （P<0.01）. Compared with model group， the p-Akt/Akt ratios in the brain tissue of the rats in medium dose and high dose of Jingkang Capsule groups were significantly increased （P<0.01 or P<0.001）， the expression levels of Bcl-2 protein were significantly increased （P<0.05 or P<0.01）， and the expression levels of Bax and Cleaved Caspase-3 proteins were significantly decreased （P<0.05 or P<0.01）. Conclusion Jingkang Capsule exerts a protective effect on the brain tissue in rats with cerebral ischemia， and its protective effect may be related to the regulation of Akt and apoptosis-related protein expression.

Objective To investigate the protective effect of human bone marrow mesenchymal stem cell-derived exosomes （hBMSCs-Exo） on neurological injury after intracerebral hemorrhage （ICH） in rats， and to elucidate the regulatory mechanism of autophagy in this process. Methods ?For in vivo experiments，thirty male SD rats were randomly divided into sham operation group， ICH group， and ICH+Exo group， with 10 rats in each group. Collagenase injection method was used to establish the rat ICH model in ICH group and ICH+Exo group. The rats in sham operation group were only drilled and injected with the same volume of normal saline. At 24 h after modeling， the rats in ICH+Exo group were injected with 20 μg hBMSCs-Exo into the brain tissue， and the rats in ICH group were only injected with phosphate-buffered saline （PBS）. The modified neurological severity scores （mNSS） of the rats in various groups were evaluated on days 1， 3， 7， 14， and 21 post-surgery. At day 7 post-surgery， Nissl staining method was used to detect the numbers of Nissl-positive neurons in the brain tissue of the rats in various groups. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL）method was used to detect the percentages of TUNEL-positive cells in the brain tissue of the rats in various groups. Immunofluorescence staining method was used to observe the expressions of neuronal nuclear protein （NeuN）， glial fibrillary acidic protein （GFAP）， and ionized calcium-binding adapter molecule 1 （Iba1） proteins in the brain tissue of the rats in various groups. Immunohistochemistry method was used to detect the expressions of autophagy-related proteins including microtubule-associated protein light chain 3 （LC3）， Beclin-1， P62， and autophagy-related protein 7 （ATG7） in the brain tissue of the rats in various groups. For in vitro experiments， 1-methyl-4-phenylpyridinium ion （MPP?） was used to induce oxidative stress in SH-SY5Y cells. The cells were divided into control group， MPP? group， MPP?+Exo group， and MPP⁺+Exo+3-methyladenine （3-MA） group. Methylthiazolyldiphenyl-tetrazolium bromide（MTT） method was used to detect the cell activities in various groups. Acridine orange （AO） staining method was used to evaluate the levels of autophagy of the cells in various groups. Results The mNSS score results showed that compared with sham operation group， the mNSS scores of the rats in ICH group were significantly increased at all time points（P<0.05）；compared with ICH group，the mNSS scores of the rats in ICH+Exo group were significantly decreased at all time points （P<0.05）. The Nissl staining results showed that compared with sham operation group，the number of Nissl-positive neurons in the brain tissue of the rats in ICH group was significantly decreased （P<0.001）；compared with ICH group，the number of Nissl-positive neurons in the brain tissue of the rats in ICH+Exo group was significantly increased （P<0.01）. The TUNEL staining results showed that compared with sham operation group，the percentage of TUNEL-positive cells in the brain tissue of the rats in ICH group was significantly increased （P<0.05） ；compared with ICH group，the percentage of TUNEL-positive cells in the brain tissue of the rats in ICH+Exo group was significantly decreased （P<0.05）. The immunofluorescence staining results showed that compared with sham operation group，the number of NeuN-positive cells in the brain tissue of the rats in ICH group was significantly decreased （P<0.05），and no statistically significant difference was observed in the percentages of positive area of GFAP and Iba1 in the brain tissue （P>0.05） ；compared with ICH group，the number of NeuN-positive cells in the brain tissue of the rats in ICH+Exo group was significantly increased （P<0.05）. The immunohistochemistry method results showed that compared with sham operation group，the percentages of positive area of LC3，Beclin-1，P62，and ATG7 proteins in the brain tissue of the rats in ICH group were significantly increased （P<0.05）；compared with ICH group，the percentage of positive area of P62 protein in the brain tissue of the rats in ICH+Exo group was significantly decreased （P<0.05）， while the percentages of positive area of LC3 and ATG7 proteins in the brain tissue of the rats were significantly increased （P<0.05 or P<0.01）. The in vitro MTT assay results showed that compared with MPP? group，the cell activity of SH-SY5Y cells in MPP?+Exo group was significantly increased （P<0.05）； compared with MPP?+Exo group， the cell activity of SH-SY5Y cells in MPP?+Exo+3-MA group was significantly decreased （P<0.05）. The AO staining method results showed that compared with MPP? group， the autophagy level of SH-SY5Y cells in MPP?+Exo group was significantly increased （P<0.05）； compared with MPP?+Exo group， the autophagy level of SH-SY5Y cells in MPP?+Exo+3-MA group was significantly decreased （P<0.05）. Conclusion ?hBMSCs-Exo alleviates neurological injury after ICH and improves neurological function in rats， and its effects are associated with promoting autophagic flux and inhibiting neuronal apoptosis. Autophagy regulation may be a key mechanism underlying its neuroprotective effect.

Objective To investigate the ameliorative effect of phloretin on pulmonary vascular remodeling in the rats with monocrotaline （MCT）-induced pulmonary arterial hypertension （PAH） and its effects on the proliferation and migration of pulmonary artery smooth muscle cells （PASMCs）， and to provide evidence for targeted therapy of PAH. Methods For in vivo experiments， twenty male Sprague-Dawley rats were randomly divided into control group， MCT group （60 mg·kg^-1）， low dose of phloretin group （10 mg·kg^-1）， and high dose of phloretin group （20 mg·kg^-1）. Echocardiography method was used to detect the pulmonary artery acceleration time （PAT） and mean pulmonary artery pressure （mPAP） in the rats in various groups. HE staining method was used to detect the percentages of pulmonary artery wall thickness （WT%） and percentages of pulmonary artery wall area （WA%） in the rats in various groups. Immunofluorescence staining method was used to detect the expression levels of proliferating cell nuclear antigen （PCNA） protein in the lung tissue of the rats in various groups. Western blotting method was used to detect the expression level of osteopontin （OPN） in the lung tissue of the rats in various groups. For in vitro experiments， the primary PASMCs were extracted from the rats using enzymatic digestion method. The cells were divided into control group， epidermal growth factor （EGF） group （20 ng·L^-1）， EGF+10 μmol·L^-1 phloretin group， and EGF+20 μmol·L^-1 phloretin group. Cell counting kit-8 （CCK-8） method was used to detect the activities of the cells in various groups. Wound healing assay and Transwell chamber assay were used to detect the wound healing rates and the numbers of migration cells of PASMCs in cells in various groups. Western blotting method was used to detect the expression levels of PCNA protein in PASMCs in the cells in various groups. Results The in vivo experiments results showed that compared with control group， PAT in the rats in MCT group was significantly shortened （P<0.05）， mPAP was significantly increased （P<0.05）， the WT% and WA% of the pulmonary artery were significantly increased （P<0.05 or P<0.01）， the expression level of PCNA protein in the lung tissue was significantly increased （P<0.05）， and the expression level of OPN protein was significantly increased （P<0.05）. Compared with MCT group， PATs in the rats in low dose of phloretin group and high dose of phloretin group were significantly prolonged （P<0.05 or P<0.01）， mPAPs were significantly decreased （P<0.05 or P<0.01）， the WT% and WA% of the pulmonary artery of the rats were significantly decreased （P<0.05 or P<0.01）， and the expression levels of PCNA protein in the lung tissue were significantly decreased （P<0.05）； the expression level of OPN protein in the lung tissue of the rats in high dose of phloretin group was significantly decreased （P<0.05）. The in vitro results showed that compared with EGF group， the cell activity of PASMCs in EGF+10 μmol·L^-1 phloretin group was significantly decreased （P<0.05）， the number of migration cells was significantly decreased （P<0.05）， the expression level of PCNA protein in the cells was significantly decreased （P<0.05）， while no significant difference was observed in the wound healing rate（P>0.05）； compared with EGF group， the cell activity of PASMCs in EGF+20 μmol·L^-1 phloretin group was significantly decreased （P<0.05）， the wound healing rate was significantly decreased （P<0.01）， the number of migration cells was significantly decreased （P<0.01）， and the expression level of PCNA protein was significantly decreased （P<0.01）. Conclusion Phloretin not only effectively ameliorates MCT-induced pulmonary vascular remodeling in the MCT-induced PAH rats， but also inhibits EGF-induced proliferation and migration of PASMCs， suggesting that phloretin may serve as a potential candidate drug for the treatment of PAH.

Objective To investigate the effect of omalizumab on the proliferation and migration of the airway smooth muscle cells （ASMCs） in the asthmatic mice by regulating wingless-type MMTV integration site family（Wnt）/β-catenin signal pathway. Methods Twelve mice were randomly divided into normal group and model group. The mouse model of asthma was constructed by sensitizing and stimulating the mixture of ovalbumin and aluminum hydroxide. The primary ASMCs were isolated from the bronchial tissue of the mice in normal group and model group. Immunofluorescence staining method was used to detect the positive expression of alpha-smooth muscle actin （α-SMA） in ASMCs. The ASMCs were divided into control group， asthma group， omalizumab group， iCRT3 （Wnt/β-catenin signaling pathway inhibitor） group， and omalizumab+SKL2001 （Wnt/β-catenin pathway activator） group. The ASMCs in control group were isolated from the bronchial tissues of mice in normal group. The ASMCs in other groups were isolated from the bronchial tissues of mice in model group and received corresponding treatments. 5-ethynyl-2'-deoxyuridine （EdU） staining method and cell counting kit-8 （CCK-8） method were used to detect the percentages of EdU-positive cells and proliferation activities in ASMCs in various groups. Wound healing assay was used to detect the wound healing rates in ASMCs in various groups. Enzyme-linked immunosorbent assay （ELISA） method was used to detect the levels of interleukin （IL）-6， IL-17， tumor necrosis factor-α （TNF-α）， IL-33， and IL-13 in the supernatants of ASMCs in various groups. Western blotting method was used to detect the expression levels of proliferating cell nuclear antigen （PCNA）， matrix metalloproteinase 9 （MMP-9）， Wnt 3a， and β-catenin proteins in ASMCs in various groups. Results Over 98% of the cells isolated from the bronchial tissues of the mice in normal group and model group exhibited positive α-SMA expression， confirming the successful isolation of ASMCs. Compared with control group， the EdU-positive rate in ASMCs， proliferation activity， wound healing rate， the levels of IL-6， IL-17， TNF-α， IL-33， and IL-13 in the supernatant， and the expression levels of PCNA， MMP-9， Wnt 3a， and β-catenin proteins in ASMCs in asthma group were significantly increased （P<0.05）. Compared with asthma group， the EdU-positive rates in ASMCs， proliferation activities， wound healing rates， the levels of IL-6， IL-17， TNF-α， IL-33， and IL-13 in the supernatants， and the expression levels of PCNA， MMP-9， Wnt 3a， and β-catenin proteins in ASMCs in omalizumab group and iCRT3 group were significantly decreased （P<0.05）. Compared with omalizumab group， the above indices in ASMCs were significantly increased in omalizumab+SKL2001 group （P<0.05）. Conclusion Omalizumab inhibits the proliferation， migration and inflammatory response of ASMCs in the asthmatic mice， and its mechanism is related to inhibiting Wnt/β-catenin signal pathway.

Objective To discuss the effects of aurone on osteoblast differentiation and its ameliorative effect on glucocorticoid-induced osteoporosis （GIOP） in rats， and to clarify its mechanism. Methods For in vitro experiments， MC3T3-E1 osteoblasts were selected. CCK-8 method was used to detect the cell activities in various groups after treatment with different concentrations of dexamethasone （Dex） and aurone to determine the optimal treatment concentrations of Dex and aurone. MC3T3-E1 osteoblasts were induced with 1 μmol·L^-1 Dex to establish a cell injury model. The cells were divided into control group， Dex group， Dex+aurone group， Dex+GNF351 group， and Dex+aurone+GNF351 group. Kit method was used to detect the level of malondialdehyde （MDA） and the activity of superoxide dismutase （SOD） in the cells in various groups； 2'，7'-dichlorofluorescein diacetate （DCFH-DA） fluorescence method was used to detect the level of reactive oxygen species （ROS） in the cells in various groups； enzyme-linked immunosorbent assay （ELISA） method was used to detect the levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-6 （IL-6） in the supernatants of the cells in various groups； alkaline phosphatase （ALP） kit method was used to detect the ALP activity in the cells in various groups； Western blotting method was used to detect the expression levels of Runt-related transcription factor 2 （RUNX-2）， osteocalcin （OCN）， collagen type Ⅰ alpha 1 chain （COL1A1）， osteopontin （OPN）， aromatic hydrocarbon receptor （AHR）， nuclear factor erythroid 2-related factor 2 （NRF2）， and NAD（P）H： quinone oxidoreductase 1 （NQO1） proteins in the cells in various groups. For in vivo experiments， 20 female rats were selected and randomly divided into sham operation group， GIOP group， GIOP+low dose of aurone （7 mg·kg^-1） group， and GIOP+high dose of aurone （14 mg·kg^-1） group， with 5 rats in each group. Except for sham operation group， the rats in various groups were intragastrically administered with 1 mg·kg^-1 Dex to establish GIOP model. The rats in various groups were intragastrically administered with drugs from 2 h after modeling； after continuous intervention for 7 weeks， blood was collected and the rats were sacrificed， and femoral tissue was collected. HE staining method was used to observe the pathological morphology of femoral tissue of the rats in various groups. Histomorphometry method was used to detect the bone mineral density （BMD） of the femoral tissue and trabecular bone volume fraction （BV/TV）， trabecular thickness （Tb.Th）， trabecular number （Tb.N）， and trabecular separation （Tb.Sp） of the rats in various groups. Western blotting method was used to detect the expression levels of AHR， NRF2， and NQO1 proteins in the femoral tissue of the rats in various groups. Results Compared with 0 μmol·L^-1 Dex group， the cell activity of MC3T3-E1 cells in 1 μmol·L^-1 Dex group was significantly decreased （P<0.01）， and 1 μmol·L^-1 Dex was determined for subsequent cell treatment. Compared with Dex group， the cell activity of MC3T3-E1 cells in Dex+5 μmol·L^-1 aurone group was significantly increased （P<0.01）， and 5 μmol·L^-1 aurone was used for subsequent cell treatment. Compared with control group， the cell activity of MC3T3-E1 cells in Dex group was significantly decreased （P<0.01）； the levels of MDA and ROS in the cells and the levels of TNF-α， IL-1β， and IL-6 in the supernatants were significantly increased （P<0.01）； the SOD activity was significantly decreased （P<0.01）； the ALP activity in the cells was significantly decreased （P<0.01）； the expression levels of RUNX-2， OCN， COL1A1， and OPN proteins in the cells were significantly decreased （P<0.01）； and the expression levels of AHR and NRF2 proteins in the nucleus and the expression level of NQO1 protein in the cells were significantly decreased （P<0.01）. Compared with Dex group， the cell activity of MC3T3-E1 cells in Dex+aurone group was significantly increased （P<0.01）； the levels of MDA and ROS in the cells and the levels of TNF-α， IL-1β， and IL-6 in the supernatants were significantly decreased （P<0.01）； the SOD activity was significantly increased （P<0.01）； the ALP activity in the cells was significantly increased （P<0.01）； and the expression levels of RUNX-2， OCN， COL1A1， and OPN proteins in the cells were significantly increased （P<0.01）. Compared with Dex+aurone group， the ALP activity in the cells in Dex+aurone+GNF351 group was significantly decreased （P<0.01）， and the expression levels of AHR and NRF2 proteins in the nucleus as well as the expression levels of NQO1， RUNX-2， and OCN proteins in the cells were significantly decreased （P<0.01）. The in vivo experiment results showed that compared with sham operation group， the trabecular bones in the femoral tissue of the rats in GIOP group were thinned， reduced in number， and diminished in area， the BMD of the femoral tissue， trabecular BV/TV， Tb.Th， and Tb.N of the rats were significantly decreased （P<0.01）， the Tb.Sp was significantly increased （P<0.01）， and the expression levels of AHR， NRF2， and NQO1 proteins in the femoral tissue of the rats were significantly decreased （P<0.01）. Compared with GIOP group， the pathological damage in femoral tissue of the rats in GIOP+low dose of aurone group and GIOP+high dose of aurone group was alleviated， the BMD of the femoral tissue， trabecular BV/TV， Tb.N， and Tb.Th were significantly increased （P<0.05）， the Tb.Sp was significantly decreased （P<0.05）， and the expression levels of AHR， NRF2， and NQO1 proteins in the femoral tissue were significantly increased （P<0.05 or P<0.01）. Conclusion Aurone promotes osteoblast differentiation and alleviates GIOP in rats， and its mechanism is potentially associated with promoting the nuclear translocation of AHR and activating the AHR/NRF2/NQO1 signaling pathway.

Objective To systematically evaluate the clinical efficacy and safety of external application of traditional Chinese medicine in the treatment of malignant pleural effusion （MPE）， and to provide evidence-based support for clinical application. Methods The databases including CNKI， VIP， Wanfang Data， Chinese Biology Medicine， PubMed， Embase， Cochrane Library， and Web of Science were searched to retrieve the studies of randomized controlled trials （RCTs） on external application of traditional Chinese medicine for MPE published from inception to March 18， 2026. The patients were divided into experimental group （receiving external application of traditional Chinese medicine alone or as an adjunct to conventional Western medical treatment） and control group （receiving conventional Western medical treatment alone or placebo external application）. RevMan 5.4 and R 4.3.3 softwares were used to conduct the meta-analysis. The outcome measures included pleural effusion response rate， Karnofsky performance status （KPS） score， KPS score improvement rate， and incidence of adverse reactions. This systematic review was prospectively registered in PROSPERO （registration number： CRD420251178277）. Results A total of 25 RCTs involving 1 616 patients were included. The meta-analysis results showed that compared with control group， the pleural effusion response rate of the patients in experimental group was significantly increased ［risk ratio （RR）=1.30， 95% confidence interval （CI）： 1.22-1.40， P<0.000 1］， the KPS score was significantly increased ［mean difference （MD）=9.28， 95%CI： 6.79-12.76， P<0.000 1］， and the KPS score improvement rate was significantly increased （RR=1.42， 95%CI： 1.15-1.74， P=0.000 9）. For safety outcomes， compared with control group， the incidences of gastrointestinal reactions （RR=0.65， 95%CI： 0.51-0.82， P=0.000 3）， leukopenia （RR=0.56， 95%CI： 0.33-0.93， P=0.03）， and chest pain （RR=0.80， 95%CI： 0.67-0.97， P=0.02） of the patients in experimental group were significantly decreased， while the incidence of local skin reactions was significantly increased （RR=8.38， 95%CI： 2.58-27.15， P=0.000 4）. Conclusion External application of traditional Chinese medicine， as an adjunctive treatment， shows potential value in improving the pleural effusion response rate and functional status in the patients with MPE， but skin adverse reactions should be closely monitored.

Objective To establish a prognostic model based on differentially expressed genes （DEGs） and differentially expressed long non-coding RNAs（DELs） of the recurrent glioblastoma （GBM） patients using the Chinese Glioma Genome Atlas （CGGA） database， to discuss the relationships between the DEGs， DELs and clinicopathological features， and to verify the findings through experiments. Methods The transcriptome data and corresponding clinical data of GBM patients， including 203 patients with primary GBM （primary GBM group） and 103 patients with recurrent GBM （recurrent GBM group）， were downloaded from the CGGA database. A Perl script was used to merge the expression profiles across all samples into a matrix， which was annotated using the Ensembl database. R packages edgeR and DESeq2 were used to filter out low-expression transcripts and to extract and normalize the data. The expression differences in transcripts between primary GBM group and recurrent GBM group were compared， the fold change（FC） was calculated， and the DEGs and DELs were screened with the criteria of |log₁₀FC|>0.5 and P<0.05. A univariate Cox proportional hazards regression model was used to analyze the relationship between DEGs and DELs and the prognosis of GBM patients. The DEGs and DELs associated with prognosis were selected to construct a multivariate Cox proportional hazards regression model， and the risk scores were calculated for all patients. Based on the median risk score， the patients were divided into low-risk and high-risk groups. The Kaplan-Meier survival analysis and receiver operating characteristic （ROC） curve were used to evaluate the predictive efficacy of the risk score for overall survival （OS）in the patients， and the differences in risk scores in the patients with different clinicopathological features were analyzed and compared. Lasso regression analysis was used to evaluate the predictive efficacies of the prognosis-related DEGs and DELs for patient survival， and a nomogram was constructed. The tumor tissue samples from 6 primary GBM patients and 6 recurrent GBM patients were selected as primary group and recurrent group， respectively. Real-time fluorescence quantitative PCR （RT-qPCR） method and fluorescence in situ hybridization （FISH） method were used to detect the expression levels of RP11-283I3.2 and the numbers of positive cells in tumor tissues of the patients in two groups. Results A total of 151 DEGs and DELs were identified between patients in primary GBM groupand recurrent GBM group，among which 12 were downregulated and 139 were upregulated. Univariate Cox regression analysis identified that 4 DEGs （CHRNA1， HIST1H4I， SLC18A3， and TMEM61） and 8 DELs （AC002454.1， AC006262.6， AC123886.2，CDKN2B-AS1，FAM225B，HAR1A，LBX1-AS1，and RP11-283I3.2） were significantly associated with the prognosis of GBM patients.The Kaplan-Meier survival analysis results showed that the OS of GBM patients in high-risk group was significantly lower than that in low-risk group （P<0.001）. The time-dependent ROC curve results showed that the areas under the curve （AUCs） of the risk score for predicting 1-， 3-， and 5-year survival probability were 0.675， 0.750， and 0.772， respectively. The risk score of GBM patients with wild-type isocitrate dehydrogenase （IDH） was significantly higher than that of patients with mutant IDH （P<0.001）. The Lasso regression analysis results indicated that RP11-283I3.2 had the highest score among the prognosis-related DEGs and DELs. Based on the expression profile， the patients were divided into Cluster 1 group and Cluster 2 group. Compared with Cluster 1 group， the patients in Cluster 2 group had lower OS （P<0.01）. The RT-qPCR method and FISH method results showed that compared with primary group， the expression level of RP11-283I3.2 and the number of positive cells in tumor tissue of the patients in recurrent group were significantly increased （P<0.001）. Conclusion A total of 12 DEGs and DELs associated with the prognosis of GBM patients were identified， and a prognostic model with good predictive efficacy was constructed. The high expression of RP11-283I3.2 in tumor tissues in recurrent GBM patients suggests its potential as a molecular biomarker for GBM recurrence.

Objective To investigate the effect of high-voltage long-duration pulsed radiofrequency （HL-PRF） on the cognitive function in patients with cervicogenic headache （CEH）， and to evaluate the potential clinical value of this therapeutic modality in improving cognitive function in the CEH patients. Methods A total of 23 patients diagnosed with CEH were enrolled. All patients underwent HL-PRF therapy targeting the affected C2 dorsal root ganglion. Before treatment and at 3 months after treatment， Beijing version of Montreal Cognitive Assessment （MoCA-BJ） was used to assess the global cognitive function of the patients. Auditory Verbal Learning Test-Huashan version （AVLT-H） subscale scores （N1， N2， N3， N4， N5， N6） were used to assess the episodic memory activities of the patients. Digit span test （DST） was used to assess the attention and working memory activities of the patients. Numerical rating scale （NRS） was used to assess the pain intensities of the patients. Visual analogue scale （VAS） for dizziness was used to assess the dizziness severities of the patients. Doppler ultrasonography was used to record the blood flow parameters of the ipsilateral C1 segment vertebral artery of the patients before treatment， immediately after treatment， and at 1 week， 1 month， and 3 months after treatment， including time-averaged mean velocity （Vm）， peak systolic velocity （PSV）， minimum velocity （Vmin）， resistance index （RI）， and pulsatility index （PI）. Pearson correlation coefficient analysis or Spearman correlation coefficient analysis was used to evaluate the correlations between the changes in pain and dizziness scores and the changes in cognitive test scores of the patients. Results The MoCA-BJ scores and DST scores at 3 months after treatment in the patients showed no statistically significant differences compared with those before treatment （P>0.05）， while all AVLT-H subscale scores were significantly increased in the patients （P<0.05）. The NRS score at 3 months after treatment was significantly decreased in the patients compared with that before treatment （P<0.001）， while VAS score for dizziness showed no statistically significant difference （P>0.05）. The Vm， PSV， and Vmin of the ipsilateral C1 segment vertebral artery of the patients immediately after treatment were significantly increased compared with those before treatment （P<0.05 or P<0.001）， while the PI and RI showed no statistically significant differences （P>0.05）. No statistically significant differences were observed in the above blood flow parameters of the patients at 1 week， 1 month， and 3 months after treatment compared with those before treatment （P>0.05）. Correlation analysis results showed that the change in VAS score for dizziness was negatively correlated with the change in AVLT-H subscale N3 score （r=-0.466， P<0.05）. Conclusion HL-PRF effectively alleviates the pain in the CEH patients， improves their performance in specific cognitive domains （e.g.， episodic memory）， and induces transient improvements in vertebral artery hemodynamics in the early period after treatment. The relief of dizziness symptoms is correlated with the improvement in certain memory function indicators.

Objective To systematically analyze the effect of antimicrobial photodynamic therapy （aPDT） on root canal microbiota in infected root canals using 16S ribosomal DNA（rDNA） amplicon sequencing technology， and to evaluate its clinical efficacy as an adjunct to root canal disinfection. Methods Fifteen patients with chronic pulpitis （CP） were enrolled as CP group， and 15 patients with chronic apical periodontitis （CAP） were enrolled as CAP group. Root canal microbiota samples were collected from patients in two groups at four key treatment stages： after access cavity preparation （Pre group）， after routine root canal preparation and disinfection （Post group）， after aPDT intervention （PDT group）， and after calcium hydroxide（CHM） dressing （CHM group）. 16S rDNA amplicon sequencing method was used to analyze qualified samples that passed quality inspection and library construction， including Alpha diversity indices （Chao1 index， Shannon index， Simpson index）， Beta diversity， and community composition differences of root canal microbiota in samples from patients in two groups. Results In samples of patients in CP group， compared with Post group， the sequencing failure rate was significantly increased in PDT group （P<0.05）， and 6 samples were subsequently included for analysis. The Alpha diversity indices in four groups of CP patients all showed a decreasing trend， with no statistically significant differences （P>0.05）. No statistically significant difference was observed in Beta diversity between Post group and PDT group of CP patients （P>0.05）. Compared with other three groups， the relative abundances of Firmicutes， Lactobacillus gasseri， and uncultured Lactobacillus were significantly increased in samples in CHM group （P<0.05）. As for samples of patients in CAP group， no statistically significant difference was observed in the sequencing failure rate between PDT group and Post group （P>0.05）， and 12 samples were subsequently included for analysis. The Chao1 index in four groups of CAP patients showed a decreasing trend， with no statistically significant differences in any Alpha diversity indices （P>0.05）. A statistically significant difference was observed in Beta diversity between post group and PDT group of CAP patients （P<0.05）. Compared with Post group， the relative abundances of Actinobacteria， Streptococcus， Actinomyces， and Porphyromonas gingivalis were significantly decreased in samples in PDT group （P<0.05）. Compared with other three groups， the relative abundances of Porphyromonas and Porphyromonas gingivalis were significantly increased in samples in CHM group （P<0.05）. Conclusion As an adjunct to root canal disinfection， aPDT exerts significant synergistic antimicrobial effects， with efficacy varying by disease type. In CP patients， aPDT enhances sterilization efficacy and shows a trend of decreasing the richness and evenness of root canal microbiota. In CAP patients， aPDT not only shows a trend of reducing root canal microbiota richness but also alters the community composition of root canal microbiota. The effect of aPDT on microbiota alterations may be associated with patient prognosis.

Objective To investigate the changes of neutrophil-to-lymphocyte ratio （NLR）， platelet-to-lymphocyte ratio （PLR）， and red cell distribution width （RDW） in the peripheral blood of the patients with post-traumatic sepsis， and to clarify their predictive values for prognosis. Methods The clinical data of 651 patients admitted to the intensive care unit （ICU） due to trauma from June 2021 to June 2025 were retrospectively analyzed. The patients were divided into sepsis group and non-sepsis group according to whether sepsis occurred during treatment. The patients in sepsis group were further divided into death group and survival group regarding their survival within 28 days after confirmed diagnosis of sepsis. The clinical indicators in the patients in various groups were compared. Multivariate Logistic regression model was used to analyze the risk factors for death in the sepsis patients. Receiver operating characteristic （ROC） curve method was used to evaluate the predictive values of the indexes for the prognosis of sepsis patients. Results A total of 436 cases （66.97%） were included in non-sepsis group， and 215 cases （33.03%） were included in sepsis group. Compared with non-sepsis group， the NLR， PLR and RDW of the patients in sepsis group were significantly increased（P<0.05）. In the patients with sepsis， 128 cases （59.53%） were included in survival group， and 87 cases （40.47%） were included in death group. Compared with survival group， the Sequential Organ Failure Assessment （SOFA） score， Acute Physiology and Chronic Health Evaluation Ⅱ （APACHE Ⅱ） score， white blood cell count （WBC）， lactate （LAC） level， NLR， PLR and RDW were significantly increased in the patients in death group （P<0.001）. Elevated SOFA score， elevated APACHE Ⅱ score， elevated LAC， elevated NLR， elevated PLR， and elevated RDW were the independent risk factors for death in sepsis patients （P<0.05）. The areas under the curve （AUCs） of LAC， NLR， PLR， RDW， combined detection of NLR， PLR and RDW， and combined detection of LAC， NLR， PLR and RDW were 0.750， 0.699， 0.740， 0.730， 0.835， and 0.875， respectively. Conclusion Elevated NLR， PLR and RDW are associated with the occurrence of post-traumatic sepsis and poor prognosis of patients. The above indices are independent risk factors for death in sepsis patients. Combined detection of NLR， PLR and RDW shows certain predictive value for the prognosis of sepsis patients.

Allogeneic bone transplantation is a commonly used method for repairing bone defects， but its long-term complications such as pathological bone resorption remain insufficiently understood in clinical practice. This article reported a rare case of progressive bone resorption occurring 2 years after allogeneic bone transplantation for calcaneal fracture， discussed its diagnostic and therapeutic strategies， and reviewed the relevant literature. The patient， a 48-year-old male， underwent open reduction and internal fixation with primary implantation of allogeneic bone graft for bilateral calcaneal fractures 7 years ago， and developed pain in both heels in the second postoperative year， which progressively worsened annually. Imaging examinations revealed extensive cavitary bone defects in both calcanei， accompanied by osteomyelitis-like changes. Intraoperative exploration identified abundant light yellow serous fluid， sandy bone debris， and granulation tissue within the calcanei. Postoperative pathological examination showed chronic inflammatory cell infiltration in the lesion， with negative bacterial culture， confirming the diagnosis of chronic aseptic osteomyelitis. After thorough debridement， the bone defect was reconstructed with autologous iliac bone grafting. The 6-month postoperative follow-up showed that the symptoms of the patient were significantly improved， and CT examination revealed favorable osseointegration in the bone-grafted area without signs of resorption. Delayed， progressive bone resorption may occur after allogeneic bone transplantation， and thorough removal of inflammatory lesions combined with autologous bone reconstruction can be an effective treatment for this complication.

Non?syndromic fused permanent teeth （NSFPT） is a dental developmental anomaly caused by non?systemic factors. Fused permanent teeth commonly occur between the maxillary central incisor and lateral incisor， while NSFPT resulting from fusion of bilateral maxillary permanent central incisors is extremely rare. This study reported a case of NSFPT， formulated an orthodontic treatment plan， and reviewed the relevant literature. The patient was a 15-year-old female whose chief complaint was irregular teeth and protruded lips. She had no history of systemic diseases. Imaging examination revealed increased sella-nasion-subspinal angle（SNA）， sella-nasion-supramental angle（SNB）， and upper incisor to sella-nasion plane angle（U1-SN） in the patient， as well as complete crown?root fusion of the bilateral maxillary permanent central incisors. The diagnoses were fused teeth of 11 and 21， Angle Class Ⅱ division 1 malocclusion， and convex facial profile. The proposed treatment plan involved extraction of the fused teeth and bilateral mandibular second premolars， followed by combined orthodontic?prosthodontic treatment to restore normal facial profile and dental arch morphology. However， the patient declined treatment due to personal reasons. For patients with fused teeth， diagnosis should be established through intraoral examination combined with imaging studies. Orthodontic treatment planning for such cases requires comprehensive consideration of multiple factors， including the degree of dental crowding and the facial profile， to determine whether extraction-based treatment is indicated. Early detection and intervention are essential to minimize the impact on the patient’s facial aesthetics and dentition， and to reduce the complexity of subsequent orthodontic treatment.

Thyroid tumors are mainly classified into epithelial and non-epithelial types， among which epithelial tumors account for 95%-99% and originate from follicular epithelial cells. Non-epithelial tumors mostly arise from lymphoid tissue， and a very small proportion originate from mesenchymal tissue， also known as mesenchymal tumors， accounting for less than 0.5%. Clear cell sarcoma of soft tissue （CCSST）， as a special type of mesenchymal tumor， is exceptionally uncommon and associated with an unfavorable prognosis. This article reported a case of primary thyroid CCSST in a patient whose tumor was confirmed by genetic testing to carry EWSR1 gene rearrangement （22q12 break）， and the relevant literatures were reviewed to summarize the clinical characteristics and experience in diagnosis and treatment. The patient， a 57-year-old female， underwent regular follow-up after radionuclide therapy for hyperthyroidism more than 10 years ago. A thyroid nodule was detected during the physical examination one year ago， and after further evaluation， she was admitted for total bilateral thyroidectomy. Postoperative paraffin pathology and immunohistochemistry showed that the tumor cells were positive for S-100 and sex-determining region Y-box 10（SOX10） and negative for human melanoma black 45（HMB45）. Fluorescence in situ hybridization （FISH） confirmed the presence of EWSR1 gene rearrangement （22q12 break）， leading to a definite diagnosis of primary thyroid CCSST with EWSR1 gene rearrangement. Postoperative positron emission tomography-computed tomography （PET-CT） excluded metastasis elsewhere in the body. In clinical practice， rare genetic alterations， such as EWSR1 rearrangement， should be carefully considered in the patients with unusual thyroid tumors. Individualized diagnosis and therapeutic strategies， together with long-term follow-up schedules should be formulated to achieve early identification and precise intervention， thereby improving the long-term prognosis of patients.

Objective To investigate the safety of gallbladder-preserving cholelithotomy via natural orifice transluminal endoscopic surgery （NOTES） and the feasibility of the transgastric tunnel approach. Methods Six miniature pigs weighing 18±2 kg were selected， and gallstone models were established by feeding method. The miniature pigs with successful modeling were randomly divided into tunnel group （submucosal tunnel approach） and control group （layered gastric wall incision approach） using a random number table method， with 3 miniature pigs in each group. Both groups of miniature pigs entered the abdominal cavity through an incision in the anterior wall of the gastric antrum， underwent cholecystotomy and stone removal， and then the gallbladder and gastric wall incisions were closed with metal clips. The surgical time， blood loss， and occurrence of complications in the two groups of miniature pigs were recorded. Incision healing was observed by anatomical dissection at 1 month postoperatively. White blood cell （WBC） count， C-reactive protein （CRP） level， and total bilirubin （TBIL） level， and alanine aminotransferase （ALT） and aspartate aminotransferase （AST） activities in the serum of the miniature pigs were detected at 30 min before surgery， 24 and 72 h postoperatively， and 1 month postoperatively. Results All 6 miniature pigs successfully completed the surgery. Gallstones （1-3 stones with diameters of approximately 1-8 mm） were successfully formed in 67% （4/6） of the model miniature pigs. Compared with control group， the gastric wall incision time in tunnel group was significantly increased （P<0.05）. Compared with control group， the blood loss from gastric wall incision was less in tunnel group， but no statistically significant difference was observed （P>0.05）. Compared with control group， the time required for gastric wall closure was significantly decreased in tunnel group （P<0.05）， and the number of metal clips used was significantly decreased （P<0.05）. No serious complications occurred in the miniature pigs in two groups postoperatively， and anatomical dissection at 1 month postoperatively showed good incision healing. For laboratory indicators， compared with those at preoperative 30 min， the WBC count and CRP level in the serum of the miniature pigs were significantly increased at 24 h postoperatively （P<0.01）. At 72 h postoperatively， the WBC count in the serum of the miniature pigs returned to normal， while the CRP level remained significantly increased （P<0.01）. Compared with those at preoperative 30 min， no statistically significant differences were observed in TBIL levels or ALT and AST activities of the miniature pigs at any postoperative time point （P>0.05）. Conclusion Transgastric NOTES gallbladder-preserving cholelithotomy is safe and feasible. The tunnel approach shows advantages in reducing bleeding and facilitating gastric wall incision closure.

Objective To explore the regulatory effect of ultrasonic power on the dispersibility of graphene oxide （GO）， to reveal the correlation between GO dispersibility and the microstructure as well as biological properties of GO/zinc film composite coating materials， and to optimize the preparation process of GO-based biomedical coatings. Methods GO powder （purity≥99%） and zinc film were used as raw materials. GO dispersion systems and GO/zinc film composite coating materials were prepared by a stirring-coupled ultrasonic dispersion-dip coating process， with three ultrasonic power gradients set at 100 W， 300 W， and 600 W. Scanning electron microscope （SEM） and Raman spectroscopy were used to analyze the surface dispersion morphology， surface sheet size， coating porosity， and structural defects of GO on the GO/zinc film composite coating materials under different ultrasonic powers. Live/dead cell staining and cell counting kit-8 （CCK-8） method were used to evaluate the survival and the proliferative activities of mouse mesenchymal stem cells （MSCs） in various groups treated with GO dispersions prepared under different ultrasonic powers. Plate counting method was used to determine the survival rates of Escherichia coli （E.coli） and Staphylococcus aureus （S.aureus） in various groups after treatment with GO dispersions prepared under different ultrasonic powers. Results The SEM observation showed that the ultrasonically dispersed GO coating achieved uniform and continuous coverage on zinc film surface； the GO/zinc film coating material in 300 W group exhibited a smooth and dense surface. The surface sheet size and rate of porosity of the GO/zinc film material in 300 W group were moderate， which were lower than those in 100 W group and higher than those in 600 W group （P<0.05）. The Raman spectroscopy analysis results revealed that the ratio of intensities of defect peak and graphitic peak （I_D/I_G） of the GO/zinc film material in 300 W group was higher than that in 100 W group but lower than that in 600 W group， indicating moderate graphitization degree and structural defects. The results of live/dead cell staining showed that a high proportion of survived cells with well-spread morphology and clear pseudopodia was observed in MSCs in 300 W group； a small number of dead cells， with surviving cells exhibiting contracted cell bodies and limited spreading range was observed in the MSCs in 100 W group， while more dead cells were seen in the MSCs in 600 W group. The results of CCK-8 method demonstrated that after 108 h of intervention， compared with 100 W group and 600 W group， the proliferative activity of mouse MSCs in 300 W group was significantly increased （P<0.05）. The results of plate counting method showed that compared with 100 W group， the survival rates of E.coli and S.aureus in 300 W group were significantly decreased （P<0.05）； compared with 600 W group， the survival rate of S.aureus in 300 W group was significantly decreased （P<0.05）. Conclusion 300 W is identified as the optimal ultrasonic power for preparing GO/zinc film composite coating materials. This parameter can precisely regulate the dispersibility of GO， simultaneously optimize the interfacial bonding performance and biological activity of the coating， and synergistically improve biosafety and antibacterial performance. The findings provide the key process parameters and theoretical basis for the industrial production and clinical translation of GO-based biomedical coatings.

Glycosylation and ubiquitination are two prevalent forms of post-translational modification（PTM） of proteins. They form a dynamic regulatory network through their interactions， jointly participating in the regulation of key biological processes such as maintenance of protein homeostasis， signal transduction cascades， and cellular differentiation and development. Glycosylation can influence the ubiquitination process by regulating the activity and substrate specificity of ubiquitin ligase. Conversely， ubiquitination can reciprocally regulate the occurrence and types of glycosylation by altering the stability of glycosyltransferases and the spatial conformation of their substrates. Current evidence indicates that the dysregulation of the dynamic balance of these two modifications and the disruption of their regulatory network are closely associated with pathological processes such as tumorigenesis and progression， viral immune evasion， and metabolic syndrome. Based on recent domestic and international literature， this article summarized the biological functions of glycosylation and ubiquitination modifications， as well as their mutual regulatory roles in the development and progression of diseases including tumor and viral infection. The feasibility of targeting key molecules involved in the glycosylation and ubiquitination processes as potential therapeutic targets was also discussed， aiming to provide theoretical support for the treatment strategies of tumor and viral infection.

Insulin-like growth factor 2 mRNA binding protein 3 （IGF2BP3） is a critical RNA-binding protein in organisms， playing key roles in the regulation of mRNA localization， stability， and post-transcriptional translation. N6-methyladenosine （m⁶A） is one of the most prevalent modifications on mammalian mRNA， and IGF2BP3 may function as a reader protein for m⁶A methylation modifications involved in regulating mRNA metabolism. IGF2BP3 is highly expressed in various malignant tumors and is associated with poor prognosis in the patients. Targeting IGF2BP3 or its upstream and downstream regulatory networks may represent a novel strategy for cancer therapy. This review summarized the roles and potential mechanisms of IGF2BP3， acting as both a RNA-binding protein and an m⁶A reader protein， in regulating malignant biological behaviors including proliferation， invasion， metabolic reprogramming， and drug resistance in various tumor cells such as glioblastoma（BGM）， breast cancer（BC）， and lung cancer， in order to provide novel insights for the treatment of malignant tumors.

Deubiquitinating enzymes （DUBs） play crucial roles in maintaining cellular homeostasis and regulating virus-host interactions by modulating the ubiquitinated status of proteins. During human immunodeficiency virus （HIV） infection， DUBs participate in multiple critical stages of the viral lifecycle， including viral transcription， replication， regulation of host restriction factor stability， and immune evasion， demonstrating their antiviral potential. By regulating key HIV-related molecules， DUBs can directly or indirectly influence HIV replication and dissemination， thereby representing important potential targets for antiviral therapy. However， the DUBs family comprises numerous members with complex functions， and its regulatory network interacts with other post-translational modifications， posing challenges for drug development targeting DUBs. This article systematically reviewed the mechanisms of DUBs in HIV infection， with a focus on their regulation of host restriction factors and key viral proteins， and discussed the significance and potential value of developing novel antiviral drugs targeting DUBs. Through an in-depth analysis of the mechanism of DUBs and the integration of new drug development technologies， it is anticipated that efficient and precise DUB-targeted therapeutic strategies can be achieved， providing new insights and theoretical support for HIV treatment and interventions against other viral infections.

Pancreatic cancer is a gastrointestinal malignant tumor with insidious onset，high malignancy， and extremely poor prognosis. Its clinical treatment faces multiple challenges， such as difficulty in early diagnosis， high metastatic potential， and strong drug resistance. Long non-coding RNAs （lncRNAs） play a key role in the regulation of gene expression in pancreatic cancer through competing endogenous RNA （ceRNA） mechanisms. However， the role and mechanism of lncRNA-mediated ceRNA regulation network in the occurrence and development of pancreatic cancer have not been systematically reviewed， and there is a lack of sufficient theoretical basis for its clinical translation and application prospects. This review summarized recent research advances on the lncRNA-mediated ceRNA regulatory mechanism in the development and progression of pancreatic cancer，systematically summarizing its specific molecular mechanisms in tumor proliferation， metastasis， epithelial-mesenchymal transition（EMT）， tumor microenvironment remodeling， and chemoresistance. It focused on exploring potential therapeutic targets and the derived intervention strategies， and summarized the clinical application value and development prospects of lncRNAs as potential biomarkers. Research on lncRNA-mediated ceRNA regulatory mechanisms is expected to provide new insights for the precise diagnosis and treatment of pancreatic cancer.

The oligoadenylate synthetase （OAS） family comprises interferon（IFN）-inducible antiviral proteins that are widely involved in biological processes including viral recognition， 2'-5'-linked oligoadenylate polymers（2-5A） synthesis， ribonuclease L （RNase L） activation， and cellular apoptosis. In recent years， their roles in oncology have attracted increasing attention. OAS family members exhibit aberrant expression in various solid tumors and hematological malignancies， participating in the regulation of tumorigenesis， tumor progression， immune evasion， and therapeutic resistance， thereby holding significant research value and clinical translational potential. In solid tumors， elevated expression of OAS1， OAS2， OAS3， and OAS like protein（OASL） is closely associated with poor prognosis， immune infiltration characteristics， and chemoresistance in cancer patients. Several members have been incorporated into prognostic prediction models across multiple studies， suggesting their potential utility as prognostic biomarkers and therapeutic targets. In hematological malignancies， OAS family members also participate in immune modulation and drug sensitivity regulation. This review systematically summarizes the structure and function of the OAS gene family， with a focus on the expression profiles， mechanistic roles， and clinical significance of OAS1， OAS2， OAS3， and OASL in different tumor types. Furthermore， we discuss their application prospects as diagnostic and therapeutic targets， along with the limitations of current research， aiming to provide a reference for subsequent basic and translational investigations.