Objective To discuss the effects of sirtuin 3 （SIRT3） deficiency on the pathological damage of periodontal tissues， osteoclast activity， and macrophage polarization in the mice with periodontitis， and to clarify the molecular mechanism. Methods Twelve C57BL/6 mice were randomly divided into SIRT3 wild-type （WT） group（SIRT3^+/+ group） and SIRT3 knockout group（SIRT3^-/- group）， with 6 mice in each group. A mouse periodontitis model was established， and the samples were collected on the 7th and 14th days of modeling. HE staining， Safranin O-Fast Green staining， and tartrate-resistant acid phosphatase （TRAP） staining were used to observe the patholmorphology and osteoclast distribution in periodontal tissues of the mice in two groups； immunohistochemistry was used to detect the expression levels of inducible nitric oxide synthase （iNOS） and cluster of differentiation 163 （CD163） proteins in periodontal tissues of the mice in two groups； Western blotting method was used to detect the expression levels of matrix metalloproteinase-9 （MMP-9）， phosphorylated-P65（p-P65）， and cathepsin K （CTSK） proteins in aperiodontal tissues of the mice in two groups； real-time fluorescence quantitative PCR（RT-qPCR） method was used to detect the expression levels of acid phosphatase 5 （ACP5）， MMP-9， nuclear factor of activated T cells cytoplasmic 1 （NFATC1）， and tumor necrosis factor α （TNF-α） mRNA in periodontal tissues of the mice in two groups； Micro-CT scanning was used to analyze the changes of alveolar bone microstructure and the surface area of alveolar bone damage in periodontal tissues of the mice in two groups. Results The HE staining results showed that in SIRT3^+/+ group， the periodontal ligament fibers in periodontal tissues were arranged neatly， with less inflammatory infiltration and bone resorption lacunae； in SIRT3^-/- group， the periodontal ligament fibers in periodontal tissues were arranged disorderly， with pervodontal ligament edema and widening， a large number of neutrophils and lymphocytes diffusely infiltrated， and the alveolar bone margin showed moth-eaten defects with multinucleated osteoclasts. Compared with SIRT3^+/+ group on the 7th day of modeling， the percentage of inflammatory area in periodontal ligament region of the mice in SIRT3^-/- group was significantly increased （P<0.05）； at 14th day of modeling， compared with SIRT3^+/+ group， the percentage of inflammatory area in periodontal ligament region of the mice in SIRT3^-/- group was significantly increased （P<0.01）. The Safranin O-Fast Green staining results showed that in SIRT3^+/+ group， the bone matrix of alveolar bone was uniformly green， the trabecular surface was smooth and continuous， a few scattered red granules were observed in the bone lacunae， the boundary between bone matrix and bone marrow cavity was clear without significant staining loss， and the collagen fibers were densely and neatly arranged without signs of fracture or dissolution； in SIRT3^-/- group， the green staining of alveolar bone matrix was significantly lighter and showed patchy loss， the collagen fibers were fractured and dissolved， the trabecular surface was rough and irregular， the red safranin positive area was significantly reduced or even disappeared， and multiple unstained blank areas appeared in the bone matrix， especially around bone resorption lacunae. Compared with SIRT3^+/+ group on the 7th and 14th days of modeling， the percentages of bone matrix degradation area of the mice in SIRT3^-/- group were significantly increased （P<0.01）. The TRAP staining results showed that on the 7th day of modeling， compared with SIRT3^+/+ group， the steoclast activity in periodontal tissue of the mice in SIRT3^-/- group was significantly increased （P<0.05）； on the 14th day of modeling， compared with SIRT3^+/+ group， the steoclast activity in periodontal tissue of the mice in SIRT3^-/- group was significantly increased （P<0.01）. The immunohistochemistry results showed that a large number of iNOS strongly positive cells were observed in periodontal tissue of the mice in SIRT3^-/- group， distributed in patches； while in SIRT3^+/+ group， there were few positive cells with light staining； CD163 positive cells in periodontal tissue of the mice in two groups were scattered and weakly stained. Compared with SIRT3^+/+ group， the expression levels of iNOS protein in periodontal tissue of the mice in SIRT3^-/- group was significantly increased （P<0.01）. The Western blotting results showed that compared with SIRT3^+/+ group， the expression levels of MMP-9 and p-P65 proteins in periodontal tissue of the mice in SIRT3^-/- group were significantly increased （P<0.05）， and the expression level of CTSK protein was significantly decreased （P<0.05）. The RT-qPCR method results showed that at 3rd day of modeling， compared with SIRT3^+/+ group， the expression levels of TNF-α， ACP5， MMP-9， and NFATC1 mRNA in periodontal tissues of the mice in SIRT3^-/- group were significantly increased （P<0.01）； on the 5th and 7th days of modeling， compared with SIRT3^+/+ group， the expression levels of ACP5 and MMP-9 mRNA in periodontal tissues of the mice in SIRT3^-/- group were significantly increased （P<0.01）. The Micro-CT scanning results showed that on the 14th day of periodontitis model， obvious alveolar bone resorption and relaxation and erosion of gingiva tissue were observed in the alveolar bone tissue of the mice. Compared with SIRT3^+/+ group， the surface area of alveolar bone damage of the mice in SIRT3^-/- group was significantly increased （P<0.05）. Conclusion SIRT3 deficiency aggravates alveolar bone resorption and periodontal tissue damage in a mouse periodontitis model， and the mechanism may be related to upregulation of MMP-9 and nuclear factor-kappa B（NF-κB） signaling pathways， promotion of macrophage polarization toward the pro-inflammatory M1 phenotype， and subsequent enhancement of osteoclast activity and bone matrix degradation.

Objective To discuss the effect of Pueraria lobata-derived exosome-like nanovesicles （PL-ELNs） on senescence of L929 mouse fibroblasts induced by D-galactose （D-gal）， and to clarify the possible mechanism of action. Methods The PL-ELNs were extracted from Pueraria lobata by differential centrifugation combined with sucrose density gradient centrifugation. Transmission electron microscope （TEM） was used to observe the morphology of PL-ELNs； nanoparticle tracking analysis （NTA） was used to identify the particle size of PL-ELNs； kit method was used to quantify the protein concentration of PL-ELNs. The L929 cells were divided into control group， D-gal group， and 4， 8 and 16 mg·L^-1 PL-ELNs+D-gal groups. The PKH26-labeled PL-ELNs were used to treat the L929 cells， and the uptake of PL-ELNs by L929 cells was observed. D-gal was used to establish the senescence model of L929 cells. CCK-8 method was used to screen the optimal concentrations of D-gal and PL-ELNs and to detect the activities of the cells in various groups； SA-β-gal staining was used to observe the staining of L929 cells in various groups and to calculate the percentage of SA-β-gal staining positive cells. EdU staining was used to detect the percentages of EdU positive cells in L929 cells in various groups； flow cytometry was used to detect the intracellular reactive oxygen species （ROS） levels and apoptotic rates of L929 cells in various groups； Western blotting method was used to detect the expression levels of collagen type Ⅰ （ColⅠ）， collagen type Ⅲ （ColⅢ）， P21 and P16 proteins in L929 cells in various groups. Results The PL-ELNs with a saucer-like morphology and a median particle size of 142.3 nm were isolated from Pueraria lobata， and could be effectively taken up by L929 cells. D-gal at 20 g·L?1 successfully induced senescence of L929 cells. The CCK-8 assay results showed that compared with control group， the activity of L929 cells in D-gal group was significantly decreased （P<0.01）； compared with D-gal group， the activities of L929 cells in 4， 8 and 16 mg·L^-1 PL-ELNs+D-gal groups were significantly increased （P<0.01）. The SA-β-gal staining results showed that compared with control group， the percentage of SA-β-gal staining positive L929 cells in D-gal group was significantly increased （P<0.01）； compared with D-gal group， the percentages of SA-β-gal staining positive L929 cells in 4， 8 and 16 mg·L^-1 PL-ELNs+D-gal groups were significantly decreased （P<0.05 or P<0.01）. The Western blotting results showed that compared with control group， the expression levels of P21 and P16 proteins in L929 cells in D-gal group were significantly increased （P<0.01）； compared with D-gal group， the expression levels of P21 and P16 proteins in L929 cells in 8 and 16 mg·L^-1 PL-ELNs+D-gal groups were significantly decreased （P<0.05 or P<0.01）. The EdU staining results showed that compared with control group， the percentage of EdU positive L929 cells in D-gal group was significantly decreased （P<0.01）； compared with D-gal group， the percentages of EdU positive L929 cells in 4， 8 and 16 mg·L^-1 PL-ELNs+D-gal groups were significantly increased （P<0.05 or P<0.01）. The flow cytometry results showed that compared with control group， the ROS levels in L929 cells in D-gal group was significantly increased （P<0.01）； compared with D-gal group， the ROS levels in L929 cells in 8 and 16 mg·L^-1 PL-ELNs+D-gal groups were significantly decreased （P<0.05 or P<0.01）. Compared with control group， the apoptotic rate of L929 cells in D-gal group was significantly increased （P<0.01）； compared with D-gal group， the apoptotic rates of L929 cells in 4， 8 and 16 mg·L^-1 PL-ELNs+D-gal groups were significantly decreased （P<0.01）. The Western blotting results showed that compared with control group， the expression levels of ColⅠ and ColⅢ proteins in L929 cells in D-gal group were significantly decreased （P<0.01）； compared with D-gal group， the expression levels of ColⅠ and ColⅢ proteins in L929 cells in 8 and 16 mg·L^-1 PL-ELNs+D-gal groups were significantly increased （P<0.05 or P<0.01）. Conclusion PL-ELNs can ameliorate D-gal-induced senescence of L929 cells， and the mechanism may be related to increasing the proliferation activity of senescent cells， reducing apoptosis， and decreasing the ROS level and the expression levels of P21 and P16 proteins in the cells.

Objective To discuss the effect of exosomes derived from bone marrow mesenchymal stem cells（BMSCs） （BMSC-exos） on asthma in the mice， and to clarify the mechanism. Methods The BMSCs were extracted， and the expressions of its markers were detected. The BMSCs were transfected with microRNA （miR）-26a mimic （mimic-miR-26a）， miR-NC and miR-26a inhibitor （inh-miR-26a）， respectively， and the corresponding exosomes （mimic-miR-26a exos， miR-NC exos and inh-miR-26a exos） were extracted. Mouse asthma model was established by administering ovalbumin （OVA） and aluminum hydroxide to the mice. The 50 successfully modeled mice were randomly divided into model group， BMSC-exos group， miR-NC exos group， mimic-miR-26a exos group and inh-miR-26a exos group， with 10 mice in each group， and 10 non-modeled mice were regarded as control group. Each group was administered daily with 10 μg of BMSC-exos， miR-NC exos， mimic-miR-26a exos， inh-miR-26a exos and an equal volume of phosphate buffered saline （PBS）， respectively. HE staining was used to observe the pathomorphological manifestations of lung tissue of the mice in various groups； small animal lung function test instrument was used to detect the airway resistance of mice in various groups； real-time fluorescence quantitative PCR（RT-qPCR） method was used to detect the expression levels of miR-26a in the lung tissue of mice in various groups； dual luciferase reporter gene assay was used to verify the targeting relationship between miR-26a and Toll-like receptor 4 （TLR4）； Western blotting method was used to detect the expression level of TLR4 protein in the lung tissue of mice in various groups. Results The extracted BMSCs highly expressed their markers cluster of differentiation（CD）44 and CD105， but did not express CD45 and CD34， indicating that BMSCs were successfully extracted. The particle size of BMSC-exos was about 100 nm， with a bilayer membrane and a “cup-shaped” morphology， which conformed to the structural characteristics of exosomes. BMSC-exos expressed tumor susceptibility gene 101 （TSG101）， CD9 and CD81， which conformed to the biological characteristics of exosomes. HE staining results showed that the lung tissue structure of mice in control group was normal， the alveolar septum was not significantly widened， and no obvious inflammatory cell infiltration was observed； the lung tissue structure of mice in model group was disordered， the alveolar structure was obviously damaged， the alveolar septum was significantly widened， there was a large amount of inflammatory cell infiltration， goblet cells were hyperplastic， and lung injury was obvious， indicating successful modeling； in BMSC-exos group， the lung tissue structure disorder of mice was significantly alleviated， the inflammatory cell infiltration was reduced， and the alveolar structure damage was reduced； after treatment with miR-26a BMSC-exos， the lung tissue structure of mice in control group was normal； in model group， a large amount of inflammatory cell infiltration was observed in the lung tissue， the alveolar structure was obviously damaged， goblet cells were obviously hyperplastic， the alveolar septum was significantly widened， and lung injury was obvious； compared with model group， the degree of lung tissue lesions in miR-NC exos group was alleviated， and the degree of lung tissue lesions in mimic-miR-26a exos group was further alleviated. The small animal lung function test results showed that under the action of 6.25， 12.50， 25.00 and 50.00 g·L^-1 methacholine （Mch）， compared with control group， the airway resistance of mice in model group was significantly increased （P<0.05）； compared with model group， the airway resistance of mice in BMSC-exos group was significantly decreased （P<0.05）. Under the action of 6.25， 12.50， 25.00 and 50.00 g·L^-1 Mch， compared with model group， the airway resistance of mice in miR-NC exos group was significantly decreased （P<0.05）； compared with miR-NC exos group， the airway resistance of mice in mimic-miR-26a exos group was significantly decreased （P<0.05）， and the airway resistance of mice in inh-miR-26a exos group was significantly increased （P<0.05）. The real-time fluorescence quantitative PCR （RT-qPCR） method results showed that compared with miR-NC exos， the expression level of miR-26a in mimic-miR-26a exos was significantly increased （P<0.05）， and the expression level of miR-26a in inh-miR-26a exos was significantly decreased （P<0.05）； compared with control group， the expression level of miR-26a in the lung tissue of mice in model group was significantly decreased （P<0.05）； compared with model group， the expression level of miR-26a in the lung tissue of mice in BMSC-exos group was significantly increased （P<0.05）； compared with miR-NC exos group， the expression level of miR-26a in the lung tissue of mice in mimic-miR-26a exos group was significantly increased （P<0.05）， and the expression level of miR-26a in the lung tissue of mice in inh-miR-26a exos group was significantly decreased （P<0.05）. The dual luciferase reporter gene assay results showed that compared with the cells transfected with TLR4 wild type （WT） and miR-NC group， the relative luciferase activity of cells in group transfected with TLR4 WT and miR-26a was significantly decreased （P<0.05）， indicating that TLR4 was the target gene of miR-26a. The Western blotting method results showed that compared with control group， the expression level of TLR4 protein in the lung tissue of mice in model group was significantly increased （P<0.05）； compared with model group， the expression level of TLR4 protein in the lung tissue of mice in BMSC-exos group was significantly decreased （P<0.05）； compared with miR-NC exos group， the expression level of TLR4 protein in the lung tissue of mice in mimic-miR-26a exos group was significantly decreased （P<0.05）， and the expression level of TLR4 protein in the lung tissue of mice in inh-miR-26a exos group was significantly increased （P<0.05）. Conclusion BMSC-exos can reduce airway resistance and alleviate the degree of lung tissue lesions in the asthmatic model mice， its mechanism may be related to BMSC-exos promoting the delivery of miR-26a to the lung tissue and inhibiting the expression of TLR4.

Objective To discuss the role of selenoprotein N1 （SEPN1） gene in D-galactose （D-gal）- induced senescence of L929 cells， and to clarify its signaling mechanism on cell proliferation， oxidative stress， endoplasmic reticulum stress， mitochondrial function， and extracellular matrix synthesis（ECM）. Methods D-gal was used to treat the L929 cells to establish a cell senescence model. The experiment was divided into control group and D-gal-induced group （D-gal group）. Small interfering RNA （siRNA） was used to knock down SEPN1 gene， and the cells were divided into negative control group （si-NC group） and si-SEPN1 group. SA-β-gal staining was used to observe the staining condition of L929 cells in various groups and calculate the percentage of SA-β-gal positive cells； Western blotting method was used to detect the expression levels of SEPN1， P21， P16， collagen type Ⅰ（Col Ⅰ）， collagen type Ⅲ（Col Ⅲ）， glucose-regulated protein 78 （GRP78）， and C/EBP homologous protein （CHOP） proteins in the L929 cells in various groups； 2'-7'- dichlorofluorescein diacetate （DCFH-DA） fluorescent probe method was used to detect the levels of reactive oxygen species （ROS） in the cells in various groups； kits were used to detect the mitochondrial membrane potential levels in the cells in various groups； Fluo-4 AM calcium ion （Ca²⁺） fluorescent probe was used to detect the Ca²⁺ levels in the L929 cells in various groups； 5-ethynyl-2'- deoxyuridine （EdU） staining was used to detect the proliferation activity of L929 cells in various groups； immunofluorescence method was used to detect the expression levels of Col Ⅰ and Col Ⅲ proteins in the L929 cells in various groups. Results The SA-β-gal staining results showed that compared with control group， the percentage of SA-β-gal positive cells in L929 cells in D-gal group was significantly increased （P<0.01）； compared with si-NC group， the percentage of SA-β-gal positive cells in si-SEPN1 group was significantly increased （P<0.01）. The Western blotting method results showed that compared with control group， the expression levels of P21 and P16 proteins in L929 cells in D-gal group were significantly increased （P<0.01）， and the expression level of SEPN1 protein was significantly decreased （P<0.01）. The EdU staining results showed that compared with control group， the proliferation activity of L929 cells in D-gal group was significantly decreased （P<0.01）； compared with si-NC group， the proliferation activity of L929 cells in si-SEPN1 group was significantly decreased （P<0.01）. The DCFH-DA fluorescent probe method results showed that compared with control group， the ROS level in L929 cells in D-gal group was significantly increased （P<0.01）； compared with si-NC group， the ROS level in the cells in si-SEPN1 group was significantly increased （P<0.05）. The Western blotting method results showed that compared with si-NC group， the expression levels of SEPN1， Col Ⅰ and Col Ⅲ proteins in the cells in si-SEPN1 group were significantly decreased （P<0.05 or P<0.01）， and the expression levels of P21， P16， GRP78 and CHOP proteins were significantly increased （P<0.05 or P<0.01）. The mitochondrial membrane potential assay results showed that compared with si-NC group， the level of mitochondrial membrane potential in the cells in si-SEPN1 group was significantly decreased （P<0.01）. The Fluo-4 AM calcium ion fluorescent probe results showed that compared with si-NC group， the Ca²⁺ level in the cells in si-SEPN1 group was significantly increased （P<0.05）. The immunofluorescence method results showed that compared with si-NC group， the expression levels of Col Ⅰ and Col Ⅲ proteins in the L929 cells in si-SEPN1 group were significantly decreased （P<0.05 or P<0.01）. Conclusion Knockdown of SEPN1 can promote senescence of L929 cells， and inhibit cell proliferation and synthesis of Col Ⅰ and Col Ⅲ； its mechanism may be related to induction of endoplasmic reticulum stress， impairment of mitochondrial function， Ca²⁺ overload， and increase of intracellular ROS.

Objective To discuss the effect of histone deacetylase 6 （HDAC6） inhibitor ACY-241 on macrophage polarization and the proliferation， migration and invasion of esophageal squamous cell carcinoma cells， and to clarify its mechanism. Methods Phorbol ester was used to induce human monocytic leukemia cell line THP-1 to differentiate into M0 macrophages. The M0 macrophages were co-cultured with human esophageal squamous cell carcinoma cells KYSE150 and KYSE150 cells treated with HDAC6 inhibitor ACY-241， respectively. Flow cytometry was used to detect macrophage polarization after co-culture； enzyme-linked immunosorbent assay （ELISA） was used to determine the level of CC chemokine ligand 2 （CCL2） in the supernatant of KYSE150 cells after co-culture； Western blotting method was used to detect the expression level of CC chemokine receptor 2 （CCR2） protein in macrophages in two groups after co-culture； pLVX-CCL2 lentivirus and its negative control pLVX-NC lentivirus were used to transfect KYSE150 cells， and real-time fluorescence quantitative PCR （RT-qPCR） and Western blotting methods were used to detect the transfection efficiency. The M0 macrophages were co-cultured with KYSE150 cells transfected with lentivirus and treated with ACY-241， and divided into control group， ACY-241 group， pLVX-NC+ACY-241 group and pLVX-CCL2+ACY-241 group； ELISA method was used to determine the levels of CCL2 in supernatant of KYSE150 cells in various groups； Western blotting method was used to detect the expression levels of CCR2 protein in macrophages in various groups； flow cytometry was used to detect macrophage polarization in various groups； RT-qPCR method was used to detect the expression levels of inducible nitric oxide synthase （iNOS）， interleukin-6 （IL-6）， interleukin-10 （IL-10） and arginase 1 （Arg-1） mRNA in macrophages in various groups； CCK-8 method， colony formation assay and Transwell chamber assay were used to detect the proliferation activities， colony formation numbers， number of migration cells and number of invasion cells in the KYSE150 cells in various groups， respectively. Results Compared with control group， the percentage of M1 macrophages in ACY-241 group was increased （P<0.05）， the percentage of M2 macrophages was decreased （P<0.05）， the level of CCL2 in the supernatant of KYSE150 cells was decreased （P<0.05）， and the expression level of CCR2 protein in macrophages was decreased （P<0.05）. After transfection with pLVX-CCL2 lentivirus， the expression levels of CCL2 mRNA and protein in KYSE150 cells were significantly increased （P<0.05）. Compared with control group， the expression levels of iNOS and IL-6 mRNA in macrophages in ACY-241 group were significantly increased （P<0.05）， the expression levels of IL-10 and Arg-1 mRNA were significantly decreased （P<0.05）， the proliferation activity of KYSE150 cells was decreased （P<0.05）， and the colony formation number， number of migration cells and number of invasion cells were decreased （P<0.05）. Compared with pLVX-NC+ACY-241 group， the level of CCL2 in the supernatant of KYSE150 cells in pLVX-CCL2+ACY-241 group was increased （P<0.05）， the expression level of CCR2 protein in macrophages was increased （P<0.05）， the percentage of M1 macrophages was decreased （P<0.05）， the percentage of M2 macrophages was increased （P<0.05）， the expression levels of iNOS and IL-6 mRNA in macrophages were decreased （P<0.05） and the expression levels of IL-10 and Arg-1 mRNA were increased （P<0.05）， the proliferation activity of KYSE150 cells was increased （P<0.05）， and the colony formation number， number of migration cells and number of invasion cells in KYSE150 cells were increased （P<0.05）. Conclusion HDAC6 inhibitor ACY-241 can promote macrophage polarization towards M1 type and reduce its polarization towards M2 type， and inhibit the proliferation， migration and invasion of esophageal squamous cell carcinoma KYSE150 cells； its mechanism may be related to suppressing the CCL2/CCR2 axis.

Objective To discuss the promotion effect of overexpression of circular RNA （circRNA）_ITCH on spinal cord injury （SCI） in the rats， and to clarify its possible mechanism. Methods The rat model of SCI was established by modified Allen’s method. The successfully modeled rats were randomly divided into sham operation group， SCI model group （SCI group）， empty adeno-associated virus group （AAV-NC group） and circ_ITCH overexpression adeno-associated virus group （AAV-circ_ITCH group）， with 15 rats in each group. Microsyringe was used to inject 6 μL of AAV-circ_ITCH and AAV-NC into the injured spinal cord of rats for intervention， and the rats in sham group and SCI group were injected with the same volume of normal saline. Inclined plane test and BBB locomotor rating scale were used to evaluate the hind limb motor function of rats in various groups； real-time fluorescence quantitative PCR was used to detect the expression levels of circ_ITCH in the spinal cord injury tissue of rats in various groups； HE staining was used to observe the pathomorphological manifestations of the spinal cord injury tissue of the rats in various groups； enzyme-linked immunosorbent assay was used to detect the levels of interleukin-1β （IL-1β）， interleukin-6 （IL-6） and tumor necrosis factor-α （TNF-α） in spinal cord injury tissue of the rats in various groups； Western blotting method was used to detect the expression levels of glial fibrillary acidic protein （GFAP）， neurocan and vimentin proteins in spinal cord injury tissue of the rats in various groups； immunofluorescence method was used to detect the expression of GFAP protein， a marker of astrocyte activation， in the spinal cord injury tissue of rats in various groups. Results Compared with sham operation group， the rats in SCI group showed obvious spinal cord tissue injury， the inclined plane angle and BBB score were significantly decreased （P<0.05）， the expression level of circ_ITCH in the spinal cord injury tissue was significantly decreased （P<0.05）， the levels of IL-1β， IL-6 and TNF-α were significantly increased （P<0.05）， and the expression levels of GFAP， neurocan and vimentin proteins as well as the fluorescence intensity of GFAP protein were significantly increased （P<0.05）. Compared with SCI group， the rats in AAV-circ_ITCH group showed significantly ameliorated spinal cord tissue injury， the inclined plane angle and BBB score were significantly increased （P<0.05）， the expression level of circ_ITCH in the spinal cord injury tissue was significantly increased （P<0.05）， the levels of IL-1β， IL-6 and TNF-α were significantly decreased （P<0.05）， and the expression levels of GFAP， neurocan and vimentin proteins as well as the fluorescence intensity of GFAP protein were significantly decreased （P<0.05）； in AAV-NC group， there were no statistically significant differences in the above indicators （P>0.05）. Conclusion Overexpression of circ_ITCH can alleviate spinal cord tissue injury in the rats with SCI and promote the recovery of motor function in the rats； its mechanism may be related to inhibiting neuroinflammation and astrocyte activation.

Objectives To discuss the effect of silencing X-box binding protein 1 （XBP1） on neurological functional recovery in the rats with spinal cord injury （SCI）， and to clarify its possible mechanism. Methods Seventy-five male SD rats with body mass of 200-240 g were randomly divided into sham operation group， model group （SCI group）， negative control lentivirus group （sh-NC group）， XBP1 gene silencing group （sh-XBP1 group）， and sh-XBP1+nuclear factor E2-related factor 2 （Nrf2） inhibitor ML385 group （sh-XBP1+ML385 group）， with 15 rats in each group. The modified Allen’s method was used to establish the SCI models of rats； the corresponding lentivirus were injected into the spinal cord and ML385 was intraperitoneally injected for intervention. BBB motor function score and open field test were used to evaluate the hindlimb motor function of the rats in various groups； HE staining was used to observe the histopathological morphology of spinal cord tissue of the rats in various groups； immunofluorescence method was used to observe the expression levels of neuronal specific marker neuronal nuclear protein （NeuN） in spinal cord tissue of the rats in various groups； kits was used to detect the levels of malondialdehyde （MDA） and glutathione （GSH） and the activities of superoxide dismutase （SOD） in spinal cord tissue of the rats in various groups； a kit was used to detect the levels of ferrous ion （Fe²⁺） in spinal cord tissue of the rats in various groups； Western blotting method was used to detect the expression levels of XBP1， Nrf2， glutathione peroxidase 4 （GPX4）， and solute carrier super family 7 member 11 （SLC7A11） proteins in spinal cord tissue of the rats in various groups. Results The BBB motor function score and open field test results showed that compared with sham operation group， the BBB motor function scores and total moving distances in open field test of the rats in SCI group were significantly decreased at 7， 14， 21， and 28 d after surgery （P<0.01）； compared with SCI group， the BBB motor function scores and total moving distances in open field test of the rats in sh-XBP1 group were significantly increased at 14， 21， and 28 d after surgery （P<0.05 or P<0.01）； compared with sh-XBP1 group， the BBB motor function scores and total moving distances in open field test of the rats in sh-XBP1+ML385 group were significantly decreased at 21 and 28 d after surgery （P<0.01）. The HE staining results showed that in sham operation group， the spinal cord tissue structure of the rats was intact， with uniform and clear staining； in SCI group， sh-NC group， and sh-XBP1+ML385 group， the spinal cord tissue of the rats was severely injured， with edema， disordered distribution of nerve cells， and abnormal morphology and number； in sh-XBP1 group， the spinal cord tissue structure of the rats was relatively intact， and the state of nerve cells was significantly improved. The immunofluorescence method results showed that compared with sham operation group， the expression level of NeuN protein in spinal cord tissue of the rats in SCI group was significantly decreased （P<0.01）； compared with SCI group， the expression level of NeuN protein in spinal cord tissue of the rats in sh-XBP1 group was significantly increased （P<0.01）； compared with sh-XBP1 group， the expression level of NeuN protein in spinal cord tissue of the rats in sh-XBP1+ML385 group was significantly decreased （P<0.05）. The ELISA and kit results showed that compared with sham operation group， the levels of MDA and Fe²⁺ in spinal cord tissue of the rats in SCI group were significantly increased （P<0.01）， and the level of GSH and the activity of SOD were significantly decreased （P<0.01）； compared with SCI group， the levels of MDA and Fe²⁺ in spinal cord tissue of the rats in sh-XBP1 group were significantly decreased （P<0.01）， and the level of GSH and the activity of SOD were significantly increased （P<0.01）； compared with sh-XBP1 group， the levels of MDA and Fe²⁺ in spinal cord tissue of the rats in sh-XBP1+ML385 group were significantly increased （P<0.01）， and the level of GSH and the activity of SOD were significantly decreased （P<0.01）. The Western blotting method results showed that compared with sham operation group， the expression level of XBP1 protein in spinal cord tissue of the rats in SCI group was significantly increased （P<0.01）， and the expression levels of Nrf2， GPX4， and SLC7A11 proteins were significantly decreased （P<0.01）； compared with SCI group， the expression level of XBP1 protein in spinal cord tissue of the rats in sh-XBP1 group was significantly decreased （P<0.01）， and the expression levels of Nrf2， GPX4， and SLC7A11 proteins were significantly increased （P<0.01）； compared with sh-XBP1 group， the expression levels of Nrf2， GPX4， and SLC7A11 proteins in spinal cord tissue of the rats in sh-XBP1+ML385 group were significantly decreased （P<0.01）. Conclusion Silencing of XBP1 gene can promote neurological functional recovery in the rats with SCI， and its mechanism may be related to up-regulation of Nrf2 expression and inhibition of ferroptosis in nerve cells of spinal cord tissue.

Objective To clarify the potential anti-cancer molecular targets and mechanism of grape seed proanthocyanidin extract （GSPE） on oral squamous cell carcinoma （OSCC） with network pharmacology， molecular docking technology and in vitro cell experiments， and to discuss its effects on proliferation， migration and invasion of OSCC cells. Methods Pubchem， Swisstarget Prediction， SuperPred， TargetNet， Genecards and Online Mendelian Inheriatance in Man （OMIM） databases were used to obtain the predicted targets of proanthocyanidins （PACs） and OSCC， respectively； Venn diagram was used to screen the potential target genes of PACs against OSCC； Gene Ontology （GO） functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathway enrichment analysis were performed according to David 6.8 database to predict the mechanism of action； String 12.0 was used to construct the protein-protein interaction（PPI） network； Cytoscape software and molecular docking were used to screen and preliminarily verify the binding ability of PACs with the core targets. Human tongue squamous cell carcinoma SCC-15 and CAL-27 cells were cultured in vitro， treated with different concentrations of GSPE， and divided into control group （complete medium） and 5， 10 and 20 mg·L^-1 GSPE groups. CCK-8 method was used to detect the survival rates of the cells in various groups； cell scratch assay was used to detect the scratch healing rates of the cells in various groups； Transwell chamber assay was used to detect the number of invasive cells in various groups； real-time fluorescence quantitative PCR （RT-qPCR） was used to detect the mRNA expression levels of the core targets in the cells in various groups. Results Network pharmacology predicted 50 potential targets of GSPE against OSCC. After screening and preliminary verification by molecular docking， heat shock protein 90 alpha family class A member 1（HSP90AA1）， B-cell lymphoma-2（Bcl-2）， prostaglandin-endoperoxide synthase 2（PTGS2） and nuclear factor kappa B1（NFKB1） were identified as the core targets of GSPE against OSCC. GO function enrichment analysis and KEGG signaling pathway enrichment analysis mainly involved pathways in cancer and microRNAs （miRNAs） in cancer， etc. The CCK-8 assay results showed that compared with control group， the survival rates of the cells in 5， 10 and 20 mg·L^-1 GSPE groups were significantly decreased after treatment for 12， 24， 48 and 72 h （P<0.05 or P<0.01）. The cell scratch assay results showed that compared with control group， the scratch assay rates of the cells in 5， 10 and 20 mg·L^-1 GSPE groups were significantly decreased after treatment for 12 and 24 h （P<0.05 or P<0.01）. The Transwell chamber assay results showed that compared with control group， the numbers of invasion cells in 5， 10 and 20 mg·L?1 GSPE groups were significantly decreased （P<0.01）. The RT-qPCR results showed that compared with HOK cells， the expression levels of HSP90AA1， Bcl-2， PTGS2 and NFKB1 mRNA in SCC-15 and CAL-27 cells were significantly increased （P<0.05 or P<0.01）； compared with control group， the expression levels of HSP90AA1， Bcl-2 and PTGS2 mRNA in SCC-15 cells in 5， 10 and 20 mg·L?1 GSPE groups were significantly decreased （P<0.01）， the expression levels of NFKB1 mRNA in SCC-15 cells in 10 and 20 mg·L?1 GSPE groups were significantly decreased （P<0.01）， the expression levels of HSP90AA1， Bcl-2 and NFKB1 in CAL-27 cells in 5， 10 and 20 mg·L?1 GSPE groups were significantly decreased （P<0.01）， the expression levels of PTGS2 mRNA in CAL-27 cells in 10 and 20 mg·L?1 GSPE groups were significantly decreased （P<0.01）. Conclusion GSPE exerts anti-OSCC effects by regulating pathways in cancer and miRNAs in cancer， etc. through targets such as HSP90AA1， Bcl-2， PTGS2 and NFKB1. In vitro cell experiment validation study shows that GSPE can inhibit the proliferation， migration and invasion of OSCC cells.

Objective To discuss the effect of sesamin （Ses） regulating adenosine monophosphate （AMP）-activated protein kinase （AMPK）/silent information regulator 1 （SIRT1） autophagy pathway on the learning and memory abilities of rats with Alzheimer’s disease （AD）. Methods Ninety male SD rats were randomly divided into control group， model group， low dose of Ses group （Ses-L group）， high dose of Ses group （Ses-H group）， positive control Donepezil group（Donepezil group）， and high dose of Ses+AMPK inhibitor Compound C group （Ses-H+Compound C group）， with 15 rats in each group. Step-down test was used to examine the step-down latencies and numbers of electric shocks of the rats in various groups； Morris water maze test was used to detect the escape latency and number of platform crossings of the rats in various groups； HE staining was used to observe the pathological morphology of neurons in the hippocampal CA1 region of the rats in various groups； TUNEL staining was used to detect the apoptotic rate of neurons in the hippocampus tissue of the rats in various groups； transmission electron microscope was used to observe the formation of autophagosomes in the hippocampal CA1 region of the rats in various groups； Western blotting method was used to detect the expression levels of rabbit primary antibodies against Beclin1， microtubule-associated protein 1 light chain 3 （LC3）， AMPK， phosphorylated AMPK （p-AMPK）， and SIRT1 proteins in the hippocampus CA1 region tissue of the rats in various groups. Results The step-down test， Morris water maze test and HE staining results showed that compared with control group， the step-down latency of the rats in model group was shortened （P<0.05）， and the number of electric shocks was increased （P<0.05）； the escape latency was prolonged （P<0.05）， and the number of platform crossings was decreased （P<0.05）； most neurons showed pyknosis， the number of normal cells was reduced， and the apoptotic rate of neurons was increased （P<0.05）. Compared with model group， the step-down latencies of the rats in Ses-L group， Ses-H group and Donepezil group were prolonged （P<0.05）， and the numbers of electric shocks were decreased （P<0.05）； the escape latencies were shortened （P<0.05）， and the numbers of platform crossings were increased （P<0.05）； the structure and morphology of neurons in hippocampal CA1 region were improved， and the apoptotic rates of neurons were decreased （P<0.05）. Compared with Ses-L group， the step-down latencies of the rats in Ses-H group and Donepezil group were prolonged （P<0.05）， and the number of electric shocks were decreased （P<0.05）； the escape latency was shortened （P<0.05）， and the numbers of platform crossings were increased （P<0.05）； the pathological damage in hippocampal CA1 region was further alleviated， and the apoptotic rate of neurons were decreased （P<0.05）. Compared with Ses-H group， the step-down latency of the rats in Ses-H+Compound C group was shortened （P<0.05）， and the number of electric shocks was increased （P<0.05）； the escape latency was prolonged （P<0.05）， and the number of platform crossings was decreased （P<0.05）； a large number of pyknotic neurons were observed in hippocampal CA1 region， and the apoptotic rate of neurons was increased （P<0.05）. The TUNEL staining and Western blotting results showed that compared with control group， the number of autophagosomes in hippocampal CA1 region of the rats in model group was decreased， the expression levels of Beclin1 and SIRT1 proteins and the ratios of LC3-Ⅱ/LC3-Ⅰ and p-AMPK/AMPK were decreased （P<0.05）； compared with model group， the numbers of autophagosomes in hippocampal CA1 region of the rats in Ses-L group， Ses-H group and Donepezil group were increased， the expression levels of Beclin1 and SIRT1 proteins and the ratios of LC3-Ⅱ/LC3-Ⅰ and p-AMPK/AMPK were increased （P<0.05）； compared with Ses-L group， the numbers of autophagosomes in hippocampal CA1 region of the rats in Ses-H group and Donepezil group were increased， the expression levels of Beclin1 and SIRT1 proteins and the ratios of LC3-Ⅱ/LC3-Ⅰ and p-AMPK/AMPK were increased （P<0.05）； compared with Ses-H group， the number of autophagosomes in hippocampal CA1 region of the rats in Ses-H+Compound C group was decreased， the expression levels of Beclin1 and SIRT1 proteins and the ratios of LC3-Ⅱ/LC3-Ⅰ and p-AMPK/AMPK were decreased （P<0.05）. Conclusion Ses can improve the learning and memory abilities of AD model rats， which may be related to the activation of AMPK/SIRT1 autophagy pathway by Ses.

Objective To screen the appropriate temperature and rewarming time of acute cold exposure （ACE） for establishing the rat model of myocardial injury induced by ACE， and to comprehensively evaluate the rationality and feasibility of the model establishment conditions through blood biochemistry， pathomorphology and molecular biology indicators. Methods Thirty male Wistar rats were randomly divided into -10 ℃， -15 ℃ and -20 ℃ cold exposure groups， with 10 rats in each group. The optimal modeling parameters （temperature and time） were determined by observing the mortality rates of the rats in various groups. Another fifty male Wistar rats were randomly divided into control group and rewarming （ACE/R） groups， which were further divided into 6/0， 6/6， 6/12 and 6/24 h groups according to rewarming time， with 10 rats in each group. The ACE/R model was established by cold exposure at -15 ℃ for 6 h followed by rewarming for corresponding time. The core body temperatures of the rats in various groups were detected， and the cardiac function of the rats in various groups was evaluated. HE staining was used to observe the pathological morphology of myocardium tissue of the rats in various groups； the serum levels of cardiac troponin I （cTnI） and the activities of aspartate aminotransferase （AST） and lactate dehydrogenase （LDH） of the rats in various groups were measured； enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of interleukin-1β （IL-1β）， interleukin-18 （IL-18） and interleukin-6 （IL-6） in the myocardium tissue of the rats in various groups； real-time fluorescence quantitative PCR （RT-qPCR） was used to detect the expression levels of IL-1β， IL-18， IL-6 and tumor necrosis factor-α （TNF-α） mRNA in the myocardium tissue of the rats in various groups. Results Compared with control group， the core body temperatures of the rats in ACE/R groups at different time points were significantly decreased （P<0.05 or P<0.01）. Compared with control group， the left ventricular ejection fraction （LVEF） and left ventricular fractional shortening （LVFS） of the rats in 6/6， 6/12 and 6/24 groups were significantly decreased （P<0.05 or P<0.01）， and the left ventricular internal diameter in diastole （LVIDd） and left ventricular internal diameter in systole （LVIDs） of the rats in 6/12 group were significantly increased （P<0.05）. Compared with control group， the serum level of cTnI and the activities of AST and LDH of the rats in 6/12 group were significantly increased （P<0.01）. HE staining results showed that compared with control group， the myocardium tissue of the rats in 6/12 group showed disordered arrangement of myocardial fibers， widened intercellular spaces， and a large number of inflammatory cell infiltration. The ELISA results showed that compared with control group， the levels of IL-1β， IL-18 and IL-6 in the myocardium tissue of the rats in 6/12 group were significantly increased （P<0.01）. The RT-qPCR results showed that compared with control group， the expression levels of IL-1β， IL-18， IL-6 and TNF-α mRNA in the myocardium tissue of the rats in 6/12 group were significantly increased （P<0.05 or P<0.01）. Conclusion The ACE/R myocardial injury model can be established in the male Wistar rats by exposure of cold air at -15 ℃ for 6 h followed by rewarming at room temperature for 12 h.

Objective To screen for differentially expressed genes （DEGs） between squamous cell carcinoma （abbreviated as squamous carcinoma） and adenocarcinoma based on The Cancer Genome Atlas （TCGA） database， and to investigate the role of nerve growth factor receptor （NGFR） gene in promoting apoptosis of squamous carcinoma and adenocarcinoma cells by T lymphocytes activated by programmed death receptor 1 （PD-1） inhibitor through in vitro cell experiments. Methods RNA sequencing （RNA-Seq） data of squamous carcinoma and adenocarcinoma tissues from human esophagus， lung and cervix were downloaded from TCGA database. R software DESeq2 package was used to analyze DEGs. Gene Ontology （GO） functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathway enrichment analysis were used to screen for genes related to phosphatidylinositol 3?kinase （PI3K）/protein kinase B （AKT） and Janus kinase （JAK）/signal transducer and activator of transcription （STAT） signaling pathways， and survival prognosis analysis was performed to determine NGFR gene as the outcome gene. For in vitro cell experiments， lung adenocarcinoma （LUAD） A549 cell line， cervical adenocarcinoma （CEAD） HeLa cell line， lung squamous carcinoma （LUSC） SK-MES-1 cell line and cervical squamous carcinoma （CESC） SiHa cell line were selected. The cells were divided into control group （untreated tumor cells）， OE-NC group （tumor cells 48 h after empty vector plasmid transfection）， si-NC group （tumor cells 48 h after negative siRNA transfection）， OE-NGFR group （tumor cells 48 h after NGFR gene overexpression plasmid transfection）， and si-NGFR group （tumor cells 48 h after NGFR gene siRNA transfection）. The co-culture system of tumor cells and T lymphocytes was established and divided into control group， Control+T lymphocytes group， Control+T lymphocytes+anti-PD-1 group， OE-NC/si-NC group， OE-NC/si-NC+T lymophocytes group， OE-NC/si?NC+T lymophocytes+anti-PD-1 group， OE-NGFR/si-NGFR group， OE-NGFR/si-NGFR+T lymophocytes group， and OE-NGFR/si-NGFR+T lymophocytes+anti-PD-1 group. Overexpression plasmid was used to increase the expression of NGFR gene in adenocarcinoma cells， and siRNA was used to knock down the expression of NGFR gene in squamous carcinoma cells. Real-time fluorescence quantitative PCR （RT-qPCR） was used to detect the NGFR mRNA expression level in tumor cells in various groups； Western blotting method was used to detect the NGFR protein expression level in tumor cells in various groups； flow cytometry was used to detect the apoptotic rates of tumor cells in various groups， and the relative increase folds of apoptie rates were calculated. Results The TCGA database analysis results showed that the expression level of NGFR gene in squamous carcinoma tissue was significantly higher than that in adenocarcinoma tissue （P<0.05）. The in vitro cell experiment results showed that after transfection with NGFR gene overexpression plasmid， the NGFR mRNA and protein expression levels in A549 cells and HeLa cells were significantly increased （P<0.05）； compared with Control+T lymophocytes+anti-PD-1 group and OE-NC+T lymophocytes+anti-PD-1 group， the relative increase fold in apoptotic rate of adenocarcinoma cells in OE-NGFR+T lymophocytes+anti-PD-1 group was significantly increased （P<0.05）. After transfection with NGFR gene siRNA， the NGFR mRNA and protein expression levels in SK-MES-1 cells and SiHa cells were significantly decreased （P<0.001）； compared with Control+T lymophocytes+anti-PD-1 group and si-NC+T lymophocytes+anti-PD-1 group， the relative increase fold in apoptotic rate of squamous carcinoma cells in si-NGFR+T lymophocytes+anti-PD-1 group was significantly decreased （P<0.001）. Conclusion The expression level of NGFR gene can positively regulate the pro-apoptotic effect of T lymphocytes activated by PD-1 inhibitor on the squamous carcinoma and adenocarcinoma cells. High expression of NGFR gene can enhance the sensitivity of squamous carcinoma and adenocarcinoma cells to PD-1 inhibitor， and it may serve as a potential biomarker for predicting the efficacy of PD?1 inhibitor.

Objective To investigate the effects of different degrees of cuff pressures and different intubation states on the morphology and structures of the tracheal intima in the adult rabbits， and to provide a reference for clinical tracheal intubation cuff pressure management. Methods Thirty-two New Zealand white rabbits were randomly divided into healthy state intubation group （HSI group）（n=16） and hemorrhagic shock intubation group （BLSI group）（n=16）. The hemorrhagic shock model of the rabbits in BLSI group was established. Each group was further divided into four subgroups according to cuff pressure： 30 cmH₂O （P30 cmH₂O， n=4）， 40 cmH₂O （P40 cmH₂O， n=4）， 50 cmH₂O（P50 cmH₂O， n=4）， and 60 cmH₂O（P60 cmH₂O， n=4）. The gross and pathological morphology of the tracheal intima of the rabbits at the cuff compression site in various groups were observed by visual inspection and HE staining. The various histological injury manifestations， including the rate of absence of pseudostratified ciliated columnar epithelium， the amount of inflammatory cell infiltration， the degree of submucosal congestion and edema， and submucosal hemorrhage of the rabbits in various groups were detected. Results The gross examination results revealed that the tracheal intima at the cuff compression site of the rabbits exhibited varying degrees of paleness， accompanied by peripheral bulging of the trachea； in contrast， non-compressed regions appeared dark red， and the color difference between compressed and non-compressed areas was obvious. Under the condition of the same cuff pressure， the degree of tracheal intima paleness of the rabbits in BLSI group was increased compared with HSI group. Compared with HSI group， the rates of abscence of pseudostratified ciliated columnar epithelium in the tracheal intima of the rabbits in all subgroups in BLSI group were significantly increased （P<0.05）. Compared with 30 cmH₂O pressure group， the rates of abscence of pseudostratified ciliated columnar epithelium in the tracheal intima of the rabbits in 40 cmH₂O， 50 cmH₂O， and 60 cmH₂O pressure groups were significantly elevated （P<0.05）. Compared with HSI group， the rates of inflammatory cell infiltration in the tracheal intima of the rabbits in all subgroups in BLSI group were significantly increased （P<0.05）. Compared with 30 cmH₂O pressure group， the rates of inflammatory cell infiltration in the tracheal intima of the rabbits in 40 cmH₂O， 50 cmH₂O， and 60 cmH₂O pressure groups were significantly increased （P<0.05）. Compared with HSI group， the degrees of tracheal submucosal congestion and edema of the rabbits in all subgroups in BLSI group were significantly increased （P<0.05）. Compared with 30 cmH₂O pressure group， the degrees of tracheal submucosal congestion and edema of the rabbits in 40 cmH₂O， 50 cmH₂O， and 60 cmH₂O pressure groups were significantly increased （P<0.05）.No Grade 0 injury of the rabbits was observed in either group. Conclusion Exceeding the safe upper limit of endotracheal tube cuff pressure results in varying degrees of tracheal intima injury.

Objective To analyze the active ingredient-target-signaling pathway regulatory network of Taohong Siwu Decoction （THSWD） by network pharmacology and molecular docking technology， and to verify its role in the treatment of periodontitis through in vitro experiments， and to clarify its action targets and related regulatory mechanisms. Methods The Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） was used to obtain the active ingredients and potential targets of THSWD； the Online Mendelian Inheritance in Man （OMIM）， Human Gene Database （GeneCards）， and Therapeutic Target Database （TTD） were used to screen the periodontitis-related targets； the STRING platform was used to construct a protein-protein interaction （PPI） network of the intersecting targets of THSWD and periodontitis， and to screen the core components and targets； the Metascape database was used for Gene Ontology （GO） functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathway enrichment analysis； molecular docking technology was used to verify the binding ability of the core components to the targets. In the in vitro experiments， the RAW264.7 cells were induced by Porphyromonas gingivalis lipopolysaccharide （P.g-LPS） to establish the in vitro model of periodontitis； cell counting kit-8 （CCK-8） method was used to detect the cytotoxicities of THSWD in various groups； real-time fluorescence quantitative PCR （RT-qPCR） was used to detect the mRNA expression levels of inflammatory factors in various groups； 2'，7'-dichlorodihydrofluorescein diacetate （DCFH-DA） probe method was used to detect the levels of reactive oxygen species （ROS） in the cells； Western blotting method was used to detect the expression levels of pathway-related proteins in various groups. Results Network pharmacology screening identified 45 potential active ingredients （quercetin， kaempferol， luteolin， etc.） of THSWD， and 81 intersecting genes with periodontitis， among which the core targets included prostaglandin-endoperoxide synthase 2 （PTGS2）， heat shock protein 90 alpha family class A member 1 （HSP90AA1）， B-cell lymphoma 2 （Bcl-2）， protein kinase B1 （AKT1）， mitogen-activated protein kinase 1 （MAPK1）， and v-rel reticuloendotheliosis viral oncogene homolog A （RELA）. The KEGG signaling pathway enrichment analysis results showed that THSWD might affect the inflammatory and immune processes of periodontitis by regulating signaling pathways such as the Janus kinase/signal transducer and activator of transcription （JAK/STAT） signaling pathway. The molecular docking technology results showed that the core components of THSWD had strong binding ability to the key targets of periodontitis. In the in vitro experiments， 0.2， 0.4， 0.8， and 1.6 g·L^-1 THSWD were selected as the administration concentrations， and the half-maximal inhibitory concentration （IC??） of the drug was 41.72 g·L^-1. The RT-qPCR results showed that compared with control group， the expression levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， interleukin-6 （IL-6）， and interleukin-10 （IL-10） mRNA in the cells in lipopolysaccharide （LPS） group were significantly increased （P<0.01）； compared with LPS group， the expression levels of TNF-α， IL-1β， and IL-6 mRNA in the cells in 0.2， 0.4， 0.8， and 1.6 g·L^-1 THSWD groups were significantly decreased （P<0.01）， and the expression level of IL-10 mRNA was significantly increased （P<0.05 or P<0.01）. The DCFH-DA probe method results showed that compared with control group， the ROS level in the cells in LPS group was significantly increased （P<0.01）； compared with LPS group， the ROS levels in the cells in 0.2， 0.4， and 0.8 g·L^-1 THSWD groups were significantly decreased （P<0.01）. The Western blotting method results showed that compared with control group， the expression levels of TNF-α， IL-1β， IL-6， and phosphorylated Janus kinase 1 （p-JAK1） proteins in the cells in LPS group were significantly increased （P<0.01）； compared with LPS group， the expression levels of TNF-α， IL-1β， IL-6， and p-JAK1 proteins in the cells in 0.2， 0.4， and 0.8 g·L^-1 THSWD group and ruxolitinib group were significantly decreased （P<0.05 or P<0.01）. Conclusion THSWD can treat periodontitis through multi-component， multi-target， and multi-pathway approaches.The in vitro experiments confirm that THSWD can inhibit the activation of JAK/STAT signaling pathway and has anti-inflammatory and antioxidant effects in vitro.

Objective To discuss the improvement effect of Renshen Jianzhi Tablets on cognitive function and protective effect of Penshen Jiouzhi Tablets on hippocampal neurons in cognitive impairment model rats， and to clarify its potential mechanism. Methods Forty-two SD rats were randomly divided into control group， model group， donepezil hydrochloride group （0.45 mg·kg^-1 donepezil hydrochloride）， Kaixin San group （1.35 g·kg^-1 Kaixin San）， low and high dose of Renshen Jianzhi Tablets groups （2.17 and 10.85 g·kg^-1 Renshen Jianzhi Tablets）， with 7 rats in each group. Except for control group， the rats in the other groups were subcutaneously injected with 500 mg·kg^-1 D-galactose daily to establish cognitive impairment model for 10 consecutive weeks. Morris water maze test was used to evaluate the learning and memory abilities of the rats in various groups； HE staining was used to observe the pathol morphology of hippocampus tissue of the rats in various groups. In in vitro experiments， human neuroblastoma SH-SY5Y cells were divided into control group， hydrogen peroxide （H₂O₂） group， lipopolysaccharide （LPS） group， 62.500 0， 125.000 0 and 250.000 0 mg·L^-1 Renshen Jianzhi Tablets groups. H₂O₂ and LPS were used to treat the SH-SY5Y cells to establish oxidative stress and inflammatory models， respectively. Methylthiazolyldiphenyl-tetrazolium bromide （MTT） method was used to detect the survival rate of the cells in various groups； 2'，7'-dichlorodihydrofluorescein 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 malondialdehyde （MDA）， glutathione （GSH）， tumor necrosis factor α （TNF-α）， interleukin-6 （IL-6）， interleukin-1β （IL-1β） and the activity of superoxide dismutase （SOD） in the cell supernatant in various groups. Ultra high performance liquid chromatography-tandem mass spectrometry （UHPLC-MS/MS） technology combined with chemical structure characteristics， secondary mass spectrometry fragment ion information and literature comparison was used to identify the compound components in Renshen Jianzhi Tablets. Results The Morris water maze test results showed that compared with control group， the number of platform crossings of the rats in model group was significantly decreased （P<0.01）， and the retention time in the target quadrant was significantly shortened （P<0.01）； compared with model group， the number of platform corssings of rats in Donepezil HCl group was significantly increased（P<0.01）， the number of platform crossings of the rats in low dose of Renshen Jianzhi Tablets groups was significantly increased （P<0.01）， and the retention time in the target quadrant was significantly prolonged （P<0.01）. The HE staining results showed that compared with control group， the number of hippocampal neurons of the rats in model group was decreased， the arrangement was disordered， and the intercellular space was enlarged； compared with model group， the number of hippocampal neurons of the rats in donepezil hydrochloride group， Kaixinsan group， low and high doses of Renshen Jianzhi Tablets groups was increased， the morphology was relatively regular， and the arrangement was relatively neat. The MTT assay results showed that compared with control group （P<0.01）， the survival rates of the cells in 0.625 mmol·L^-1 H₂O₂ group and 500 mg·L^-1 LPS were decreased and closest to 50%； compared with control group， reached， the cell survival rate of the cells in 500 mg·L^-1 Renshen Jianzhi Tablets group was significantly decreased （P<0.01）. The ROS fluorescence probe assay results showed that compared with control group， the ROS level in the cells in H₂O₂ group was significantly increased （P<0.01）； compared with H₂O₂ group， the ROS levels in the cells in 62.500 0， 125.000 0 and 250.000 0 mg·L^-1 Renshen Jianzhi Tablets groups were significantly decreased （P<0.01）， and the effect was most obvious in 125.000 0 mg·L^-1 Renshen Jianzhi Tablets group. The ELISA results showed that compared with control group， the MDA level in the cell supernatant in H₂O₂ group was significantly increased （P<0.01）， while the GSH level and SOD activity were significantly decreased （P<0.01）； compared with H₂O₂ group， the MDA levels in the cell supernatant in 62.500 0， 125.000 0 and 250.000 0 mg·L^-1 Renshen Jianzhi Tablets groups were significantly decreased （P<0.05 or P<0.01）， and the GSH levels and SOD activities were significantly increased （P<0.05 or P<0.01）. Compared with control group， the levels of TNF-α， IL-6 and IL-1β in the cell supernatant in LPS group were significantly increased （P<0.01）； compared with LPS group， the levels of TNF-α， IL-6 and IL-1β in the cell supernatant in 62.500 0， 125.000 0 and 250.000 0 mg·L^-1 Renshen Jianzhi Tablets groups were significantly decreased （P<0.01）. The UHPLC-MS/MS analysis results showed that a total of 73 compounds were identified in Renshen Jianzhi Tablets. Conclusion Renshen Jianzhi Tablets can effectively improve the learning and memory abilities of cognitive impairment model rats and protect the structural integrity of hippocampal neurons. Its mechanism may be related to inhibiting oxidative stress damage of nerve cells and reducing inflammatory response.

Objective To discuss the effect of Panaxatriol saponins （PTS） on D-galactose （D-gal）- induced skeletal muscle aging of mice， and to clarify its possible molecular mechanism. Methods Seventy-two male C57BL/6J mice were randomly divided into control group， D-gal group， PTS group， and D-gal+PTS group. A D-gal-induced skeletal muscle aging model was established， and then PTS was administered； the general condition of the mice in various groups was observed； the weight of rectus femoris muscle of mice was measured and the sarcopenia index （SI） was calculated； HE staining was used to observe the pathomorphology of skeletal muscle tissue of the mice in various groups； Western blotting method was used to detect the expression levels of endoplasmic reticulum（ER） stress-related proteins in the eukaryotic translation initiation factor 2α （eIF2α）/activating transcription factor 4 （ATF4） signaling pathway in skeletal muscle tissue of the mice in various groups. Results The observation of general condition showed that compared with control group， the mice in D-gal group had sparse and poor quality hair， abnormal behavior， and decreased activity； compared with D-gal group， the mice in D-gal+PTS group showed significantly improved hair condition， improved behavioral state， and were more active. The measurement results showed that compared with control group， the body mass， left rectus femoris SI， and right rectus femoris SI of the mice in D-gal group were significantly decreased （P<0.01）； compared with D-gal group， the body mass， left rectus femoris SI， and right rectus femoris SI of the mice in D-gal+PTS group were significantly increased （P<0.01）. The HE staining results showed that compared with control group， the cross-section of rectus femoris muscle of the mice in D-gal group showed broken muscle fibers， nuclear internalization of myocytes， and significantly widened intercellular space were observed， showing the morphological changes of sarcopenia； no significant differences were observed in the above tissues of the mice in PTS group； compared with D-gal group， the myocytes of the mice in D-gal+PTS group had regular morphology， no widening of intercellular space， no nuclear internalization， and the muscle morphology was significantly improved. The Western blotting method results showed that compared with control group， the ratio of phosphorylated PERK （p-PERK） to PERK （p-PERK/PERK ratio）， the expression levels of C/EBP homologous protein （CHOP） and ATF4 and activating transcription factor 5 （ATF5） proteins， and the ratio of phosphorylated eIF2α （p-eIF2α） to eIF2α （p-eIF2α/eIF2α ratio） in skeletal muscle tissue of the mice in D-gal group were significantly increased （P<0.01）； compared with D-gal group， the p-PERK/PERK ratio， p-eIF2α/eIF2α ratio， and the expression levels of ATF4 and ATF5 proteins in PTS group and D-gal+PTS group were significantly decreased （P<0.01）. The Western blotting method results also showed that compared with control group， the expression level of B-cell lymphoma 2 （Bcl-2）-associated X protein （Bax） in skeletal muscle tissue of the mice in D-gal group was significantly increased （P<0.01）， and the expression level of Bcl-2 protein was significantly decreased （P<0.01）； compared with D-gal group， the expression levels of Bax protein in skeletal muscle tissue of the mice in PTS group and D-gal+PTS group were significantly decreased （P<0.01）， and the expression levels of Bcl-2 protein in skeletal muscle tissue of the mice in PTS group and D-gal+pts group were significantly increased （P<0.01）. Conclusion PTS can ameliorate D-gal-induced skeletal muscle aging of the mice and down-regulate the expression of proteins in the eIF2α/ATF4 signal pathway； its mechanism may be related to eIF2α/ATF4 signal pathway and molecular mechaism of Bax and Bcl-2.

Objective To discuss the effect of interleukin-17 receptor B （IL-17RB） gene silencing on the proliferation， apoptosis and invasion of lung adenocarcinoma A549 cells， and to clarify its role in the occurrence and development of lung adenocarcinoma. Methods The normal human bronchial epithelial BEAS-2B cells and human lung adenocarcinoma A549 cells were cultured； real-time fluorescence quantitative PCR（RT-qPCR） and Western blotting method were used to detect the expression levels of IL-17RB mRNA and protein in the two kinds of cells. Liposome method was used to transfect IL-17RB small interfering RNA （si-RNA） fragments into the A549 cells， and the cells were divided into si-NC group and si-IL-17RB group. RT-qPCR and Western blotting methods were used to detect the silencing effect of IL-17RB； CCK-8 method was used to detect the proliferation activities of A549 cells in two groups； 5-ethynyl-2'-deoxyuridine （EdU） staining was used to detect the EdU positive cell rates of A549 cells in two groups； flow cytometry was used to detect the apoptotic rates of A549 cells in two groups； Transwell chamber assay was used to detect the numbers of migration cells and invasion cells of A549 cells in two groups； Western blotting method was used to detect the expression levels of B-cell lymphoma 2 （Bcl-2）， Bcl-2-associated X protein （Bax）， matrix metalloproteinase 9 （MMP-9）， N-cadherin， Vimentin， E-cadherin， Snail homolog （Snail） and phosphorylated P65 （p-P65） proteins in A549 cells in two groups. Results The RT-PCR results showed that compared with BEAS-2B cells， the expression levels of IL-17RB mRNA and protein in A549 cells were significantly increased （P<0.01）； compared with si-NC group， the expression levels of IL-17RB mRNA and protein in A549 cells in si-IL-17RB group were significantly decreased （P<0.01）. The CCK-8 assay results showed that compared with si-NC group， the proliferation activity of A549 cells in si-IL-17RB group was significantly decreased （P<0.01）. The EdU staining results showed that compared with si-NC group， the EdU positive cell rate of A549 cells in si-IL-17RB group was significantly decreased （P<0.01）. The flow cytometry results showed that compared with si-NC group， the apoptotic rate of A549 cells in si-IL-17RB group was significantly increased （P<0.01）. The Transwell chamber assay results showed that compared with si-NC group， the numbers of migration cells and invasion cells of A549 cells in si-IL-17RB group were significantly decreased （P<0.01）. The Western blotting results showed that compared with si-NC group， the expression levels of Bcl-2， Vimentin， N-cadherin， MMP-9， Snail and p-P65 proteins in A549 cells in si-IL-17RB group were significantly decreased （P<0.01）， while the expression levels of Bax and E-cadherin proteins were significantly increased （P<0.01）. Conclusion Silencing IL-17RB gene can reduce the proliferation and invasion abilities of lung adenocarcinoma A549 cells， promote cell apoptosis， and inhibit the expression level of p-P65 protein in cells； its mechanism may be related kinhibiting the nuclear factor-kappa B （NF-κB） singal pathway.

Objective To discuss the level of killer cell lectin-like receptor G1 （KLRG1）⁺ natural killer （NK） cells in peripheral blood of patients with non-small cell lung cancer （NSCLC）， to clarify its association with the clinical indicators， and to evaluate its clinical significance. Methods A total of 154 patients with lung adenocarcinoma who underwent surgical treatment in Sichuan Cancer Hospital from 2020 to 2022 were enrolled as the study subjects. The clinical data， such as gender， age， body mass index （BMI）， smoking history， drinking history， family history of tumor， and pathological tumor-node-metastasis （pTNM） stage， and routine blood test indicators 1-2 weeks before surgery were collected. Flow cytometry was used to detect the percentage of KLRG1⁺ NK cells in peripheral blood mononuclear cells （PBMCs）； χ2 test or Fisher’s exact test was used to analyze the association between the percentage of KLRG1⁺ NK cells and clinical characteristics； Mann-Whitney U test or Kruskal-Wallis H test was used to compare the percentages of immune cell subsets among different groups； Spearman rank correlation analysis was used to analyze the correlation between the percentage of KLRG1⁺ NK cells and routine blood test indicators. Results Among the 154 patients， there were 51 males （33.12%） and 103 females （66.88%）； 74 patients （48.05%） were younger than 55 years old， and 80 patients （51.95%） were 55 years old or older. The flow cytometry results showed that the median percentage of KLRG1⁺ NK cells in peripheral blood PBMCs was 17.05% （11.55%-23.98%）. Using this median as the cut-off value， the patients were divided into NK cell low percentage group （≤17.05%） and NK cell high percentage group （>17.05%）. The χ2 test results showed that there was a significant association between gender and the grouping of KLRG1⁺ NK cell percentage （χ2=5.78， P=0.017）， while the percentage of KLRG1⁺ NK cells was not associated with age， BMI， smoking history， drinking history， family history， or pTNM stage （P>0.05）. The median percentage of KLRG1⁺ NK cells was 54.85% （38.20%-69.00%）， and it was not associated with pTNM stage （P>0.05）. The Spearman rank correlation analysis results showed that the percentage of KLRG1⁺ NK cells was significantly positively correlated with platelet （PLT） count （rho=0.18， P=0.024）， and significantly negatively correlated with mean platelet volume （MPV） （rho=-0.24， P=0.008）. Conclusion The percentage of KLRG1⁺ NK cells in peripheral blood of patients with NSCLC has decrease trendency with tumor stage progression， and is related to PLT and MPV. The percentage of peripheral blood KLRG1⁺ NK cells combined with PLT parameters is expected to be a potential biomarker for the evaluation of NSCLC condition.

Objective To explore the characteristic metabolites in ectopic endometrium tissue of the patients with endometriosis （EMs） based on non-targeted metabolomics technology， and to clarify the key metabolic regulatory networks in the occurrence and development of the disease. Methods Paired ectopic and eutopic endometrium tissues from 91 EMs patients who（EMs group） underwent surgical treatment from January 2024 to January 2025， and normal endometrium tissue from 63 non-EMs patients（non-EMs group） were collected. Ultra-high performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） was used for non-targeted metabolomics detection and analysis. R （v4.3.0） software was used for statistical analysis of the data； principal component analysis （PCA） and orthogonal partial least squares discriminant analysis （OPLS-DA） were used to screen the characteristic metabolites of the disease； and Kyoto Encyclopedia of Genes and Genomes （KEGG） database was used for metabolic pathway enrichment analysis. Results A total of 245 endometrium tissue samples were collected， including 91 paired ectopic and eutopic endometrium tissue samples from the EMs patients and 63 normal endometrium tissue samples. The average age of patients in EMs group was （37.10±3.26） years old， and that of non-EMs group was （38.85±4.67） years old. A total of 2 116 metabolites were identified in all samples. Among the EMs patients， compared with eutopic endometrium tissue， 247 metabolites were up-regulated and 161 metabolites were down-regulated in ectopic endometrium tissue； compared with normal endometrium tissue of non-EMs patients， 412 metabolites were up-regulated and 97 metabolites were down-regulated in ectopic endometrium tissue of EMs patients； compared with normal endometrium tissue of non-EMs patients， 94 metabolites were up-regulated and 65 metabolites were down-regulated in eutopic endometrium tissue of EMs patients. Compared with both eutopic endometrium tissue of EMs patients and normal endometrium tissue of non-EMs patients， 163 metabolites represented by 6-acetamido-3-oxohexanoate were up-regulated， and 39 metabolites represented by alloxamine were down-regulated in ectopic endometrium tissue of EMs patients. KEGG metabolic pathway analysis showed that the up-regulated metabolites such as histamine， tyramine， dopamine and palmitoylethanolamide were significantly enriched in the neuroactive ligand-receptor interaction pathway （P<0.05）. Conclusion There is significant metabolic reprogramming in the ectopic endometrium tissue of EMs patients. Among them， metabolites represented by 6-acetamido-3-oxohexanoate are significantly increased in ectopic endometrium tissue， and the up-regulated metabolites are significantly enriched in the neuroactive ligand-receptor interaction pathway.

Objective To investigate the differences in the local immune microenvironment after drug-eluting bead transarterial chemoembolization （DEB-TACE） and conventional transarterial chemoembolization （c-TACE） for intermediate-advanced hepatocellular carcinoma （HCC）， and to clarify the possible mechanism by which tumor necrosis factor receptor-associated death domain （TRADD） mediates the differentiation of CD4⁺T lymphocytes into T helper 17 （Th17） lymphocytes. Methods The clinical samples from 56 patients with intermediate-advanced HCC who received TACE treatment were retrospectively collected and divided into DEB-TACE group and c-TACE group， with 28 cases in each group. High-throughput RNA sequencing was used to compare the changes in gene expression profiles and infiltration abundance of immune cells in HCC tissue before and after treatment in two groups. Flow cytometry was applied to sort CD4⁺T lymphocytes and Th17 cells from HCC tissues in two groups. The expressions of TRADD， interleukin （IL）-23A， and other related molecules in the cells were detected by real-time quantitative PCR （RT-qPCR） method， Western blotting method， and immunofluorescence method. The primary CD4⁺T lymphocytes were isolated and activated in vitro， and TRADD overexpression （TRADD-OE） and knockdown （TRADD-KD） models were constructed. The proliferation viability of CD4⁺T lymphocytes was detected by methytthiazolyl diphenyl-tetra zolilim（MTT） assay； the percentages of CD161⁺CCR6⁺ in Th17 cells in various group were detected by flow cytometry； the levels of IL-17 in the cell supernatant in various groups were measured by ELISA. The expression levels of key proteins of Janus kinase/signal transducer pathway and activator of transcription（STAT） signal phosphorylated-JAK2 （p-JAK2） and phosphorylated-STAT3 （p-STAT3） proteins and its downstream transcription factors aryl hydrocarbon receptor （AHR） and RAR-related orphan receptor gamma t （RORγt） mRNA and proteins were detected by Western blotting and RT-qPCR methods. The JAK inhibitor （Ruxolitinib） was used to intervene in TRADD-OE cells， and the aforementioned Th17 differentiation and signaling pathway indicators were detected again. Results Compared with c-TACE group， the expression levels of TRADD and IL-23A in post-operation HCC tissue in DEB-TACE group were significantly increased （P<0.05）， and the infiltration abundance of central memory CD4⁺T lymphocytes and Th17 cells was significantly increased. Compared with c-TACE group， the percentage of CD4⁺ T lymphocytes in HCC tissue in DEB-TACE group was significantly increased （P<0.05）， and the expression levels of TRADD and IL-23A mRNA and protein in the sorted CD4⁺T lymphocytes and Th17 cells were significantly increased （P<0.01）. Compared with Con-OE group， the proliferation activity of CD4⁺T lymphocytes in TRADD-OE group was significantly increased （P<0.01）； the of CD161⁺CCR6⁺ Th17 cells and the level of IL-17 in the supernatant were significantly increased （P<0.01）； the expression levels of p-JAK2， p-STAT3， AHR， and RORγt mRNA and proteins in the cells were all significantly increased （P<0.01）. Compared with Con-KD group， the above indicators in TRADD-KD group were all significantly decreased （P<0.01）. After intervention with JAK inhibitor， compared with TRADD-OE group， the percentage of CD161⁺CCR6⁺ Th17 cells and the level of IL-17 in the supernatant in TRADD-OE+JAK inhibitor group were significantly decreased （P<0.01）， and the fluorescence intensities of CD161 and CCR6， as well as the expression levels of AHR and RORγt mRNA and proteins in the cells， were significantly decreased （P<0.01）. Conclusion Compared with c-TACE， DEB-TACE can more significantly remodel the local immune microenvironment of HCC and increase Th17 cell infiltration. Its mechanism may be related to the high expression of TRADD and activating the JAK/STAT signaling pathway， thereby upregulating AHR and RORγt expressions and promoting the differentiation of CD4⁺T lymphocytes into Th17 cells.

Objective To discuss the expression of progestin and adipoQ receptor 3 （PAQR3） in hepatocellular carcinoma （HCC） and its association with immune infiltration， immunotherapy response， and patient survival， and to clarify its role in the HCC immune microenvironment and its clinical significance. Methods The HCC tissue and adjacent tissue samples were collected from 3 cases of HCC patients who underwent hepatectomy in General Hospital of Tianjin Medical University from January to December 2023. Western blotting method and immunohistochemical staining were used to detect the expression level of PAQR3 protein in HCC tissue and adjacent tissue of each HCC patient. The gene expression and clinical data of HCC patients who underwent hepatectomy were collected from The Cancer Genome Atlas （TCGA） database. Various algorithms such as CIBERSORT and TIMER2.0 were used to analyze the correlation between PAQR3 expression and immune cell infiltration； Spearman correlation analysis was used to evaluate the correlation between PAQR3 expression level and the expression levels of immune checkpoint genes ［programmed cell death protein 1 （PDCD1）， cluster of differentiation 274 （CD274）， and cytotoxic T-lymphocyte?associated protein 4 （CTLA4）］； Cox proportional hazards regression model and Kaplan-Meier method were used to analyze the relationship between PAQR3 protein expression and patient survival. Results The Western blotting and immunohistochemical staining results showed that compared with adjacent tissue， the expression level of PAQR3 protein in HCC tissue was significantly increased （P<0.05）. The TCGA database analysis results showed that compared with adjacent tissue， the expression of PAQR3 gene in HCC tissue was up?regulated （P<0.05）. The correlation analysis results showed that PAQR3 expression level was positively correlated with regulatory T cells （Tregs） （rho=0.349， P<0.001）， CD8⁺ T lymphocytes （rho=0.268， P<0.001）， M2 macrophages （rho=0.435， P<0.001）， and neutrophils （rho=0.399， P<0.001） in HCC tissue. High expression of PAQR3 protein was associated with shortened overall survival of HCC patients （P=0.019）， while immune infiltration was not associated with the survival of HCC patients （P>0.05）. The expression of PAQR3 was positively correlated with the expressions of immune checkpoint-related genes PDCD1， CD274， and CTLA4 （P<0.05）. Conclusion The expression of PAQR3 is up-regulated in HCC tissue. Its expression level is associated with the infiltration degree of various immune cells such as T lymphocytes and macrophages， the expression of immune checkpoint genes， and poor prognosis of patients， suggesting that PAQR3 may serve as a potential prognostic biomarker for HCC.

Objective To discuss the diagnostic value of differentially expressed genes （DEGs） in bone marrow mononuclear cells of the patients with acute myeloid leukemia （AML）， and to clarify their clinical significance and potential functions. Methods The GSE9476 dataset （bone marrow samples from 26 AML patients and 10 healthy donors） was obtained from the Gene Expression Omnibus （GEO） database； GEO2R was used to screen the DEGs the fold change （FC） was calculated， and |log₂ FC|≥2， P≤0.05， and adj.P≤0.05 were the criteria； the DAVID database was used to perform Gene Ontology （GO） functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathway enrichment analysis； STRING was used to construct a protein-protein interaction（PPI） network； Cytoscape was used to screen core， key， and bottleneck genes； the Venn diagram was used to obtain diagnostic candidate genes by intersection； the GEPIA and Kaplan-Meier Plotter online databases were used to test the expression differences of the candidate genes and their significance with patient survival； real-time fluorescence quantitative PCR （RT-qPCR） was used to validate the related mRNA expression levels in bone marrow mononuclear cells from clinical specimens of newly diagnosed non-acute promyelocytic leukemia （APL） type AML patients and healthy donors； through recombinant plasmid construction， lentiviral packaging and infection， AML cell models overexpressing platelet factor 4 （PF4） and pro-platelet basic protein （PPBP） were established； CCK-8 method was used to detect the proliferation activities of the cells in various groups； Western blotting method was used to detect the expression levels of apoptosis-related proteins in the cells in various groups. Results A total of 496 DEGs were screened， including 69 up-regulated genes and 427 down-regulated genes. The enrichment analysis results showed that the DEGs were mainly involved in immune response， cell cycle， and chemokine signaling pathways. The PPI network analysis identified 10 candidate genes， which were cyclin-dependent kinase 1 （CDK1）， CD36 molecule （CD36）， Runt-related transcription factor 1 （RUNX1）， CD3 delta subunit of T-cell receptor complex （CD3D）， vascular endothelial growth factor A （VEGFA）， cyclin B1 （CCNB1）， amyloid beta precursor protein （APP）， Src family tyrosine kinase （FYN）， PPBP， and PF4. The results from GEPIA and Kaplan-Meier Plotter online databases showed that compared with control group， the expression levels of APP， CD36， FYN， RUNX1， PF4， and PPBP mRNA in bone marrow aspirate fluid of the patients in AML group were increased （P<0.05）， while the expression levels of CCNB1， CD3D， CDK1， and VEGFA mRNA were decreased （P<0.05）； high expression of APP， CD36， PF4， PPBP， and RUNX1， as well as low expression of CD3D and VEGFA， were significantly associated with poor prognosis （P<0.05）. The real-time fluorescence quantitative PCR results showed that compared with control group， the expression levels of PF4 and PPBP mRNA in bone marrow aspirate fluid of the patients in AML group were significantly increased （P<0.01）. The CCK-8 assay results showed that compared with control group， after overexpression of PF4 or PPBP， the proliferation activities of THP-1 cells in OE-PF4 group and OE-PPBP group significantly increased （P<0.05）. The Western blotting method results showed that compared with control group， after overexpression of PF4 or PPBP， the expression levels of B-cell lymphoma 2 （Bcl-2） protein in THP-1 cells in OE-PF4 group and OE-PPBP group were significantly increased （P<0.05）， while the expression levels of Bcl-2-associated X protein （Bax） protein were significantly decreased （P<0.05）. Conclusion The screened PPBP and PF4 are abnormally expressed in AML and are correlated with the patient prognosis； they may participate in AML progression by promoting cell proliferation and inhibiting apoptosis， and have the potential to serve as the diagnostic and prognostic biomarkers.

Objective To discuss the risk factors in the type 2 diabetes mellitus （T2DM） patients complicated with low extremity atherosclerosis （LEAD）， and to establish a nomogram prediction model， so as to provide clinical basis for early identification of high-risk populations and formulation of intervention strategies. Methods The clinical data of 269 patients with T2DM were collected. The patients were divided into LEAD group （n=197） and non-LEAD group （n=72） according to whether they had concurrent LEAD. Univariate and multivariate Logistic regression analyses were used to analyze the risk factors of T2DM patients complicated with LEAD； the nomogram was used to construct a risk prediction model； the Bootstrap method （B=1 000） was used for internal validation； the receiver operating characteristic （ROC） curve， calibration curve， and decision curve analysis （DCA） were used to comprehensively evaluate the predictive performance of the model. Results The univariate analysis results showed that compared with non-LEAD group， the age， disease duration， fasting plasma glucose （FBG）， body mass index（BMI）， triglyceride （TG）， smoking history， and history of hypertension in the patients in LEAD group had statistically significant differences （P<0.05）. The multivariate Logistic regression analysis results showed that patients’ age ［odds ratio （OR）=1.082， 95% confidence interval （CI）： 1.039-1.127， P<0.001］， BMI（OR=1.287， 95%CI： 1.154-1.436， P<0.001）， and FBG （OR=1.159， 95%CI： 1.043-1.288， P=0.006） were the independent risk factors for T2DM complicated with LEAD. The Bootstrap internal consistency index （C-index） was 0.841 （95%CI： 0.793-0.890）， indicating that the model had good discriminative performance. The calibration curve analysis results showed that the Brier score was 0.138， and the predicted probability was highly consistent with the actual observed probability， indicating good calibration of the model. The DCA standardized net benefit value ranged from 0.2 to 0.8， indicating that the model had a significant net benefit advantage. Conclusion High BMI level is an important influencing factor for the T2DM patients complicated with LEAD. The constructed nomogram risk model based on the risk factors has good predictive performance.

The patients with herpes zoster and thoracic spine injury or degeneration often suffer from thoracic neuralgia. In severe cases， severe pain may affect sleep and daily life， and conservative treatment often yields poor results. At present， the most common minimally invasive surgical option in clinical practice is CT-guided thoracic spinal nerve root radiofrequency ablation. However， the application of CT guidance has certain limitations， such as a history of tumor in the patient， or physical inability to maintain the prone position for a long time， which may affect the operation， and the problems such as excessive radiation exposure and（or） puncture failure often occur. This article reported the application of ultrasound combined with CT-guided percutaneous spinal nerve root radiofrequency technique （PSNRRT） in the treatment of thoracic neuralgia， which effectively assisted the successful completion of the operation. Patient 1， a 63-year-old female， had postherpetic neuralgia （PHN）； Patient 2， a 58-year-old male， had lumbar vertebral fracture complicated with thoracic neuralgia after a car accident. Both patients had difficulty in maintaining the prone position for a long time， and the operations were successfully completed using ultrasound combined with CT-guided PSNRRT. The postoperative records showed that the operation time and the number of radiation exposures in both patients were less than those in patients who underwent CT-guided surgery alone， and there was no significant difference in the evaluation of treatment effect. Puncture under ultrasound combined with CT guidance is performed under real-time visualization， which may reduce radiation and shorten operation time. This method not only ensures the accuracy and safety of puncture， but also reduces operation time and radiation exposure time， and the postoperative effect is definite. This study provides a new idea for the radiofrequency treatment of thoracic spinal neuralgia， and its feasibility is further confirmed by clinical operation. The long-term efficacy and advantages need to be further verified by large-sample studies.

Primary renal angiosarcoma （RA） is clinically rare and highly aggressive， and is easily misdiagnosed at the early stage. A 70-year-old female patient admitted to our hospital with sudden severe pain in the right lower abdomen for 10 h was reported. Contrast-enhanced whole abdominal CT showed right renal rupture with active retroperitoneal hemorrhage， and emergency right nephrectomy was performed. Postoperative pathological examination confirmed moderately differentiated RA； immunohistochemical staining results showed that CD31， CD34， and ETS-related gene protein （ERG） were diffusely strongly positive， and the Ki-67 index was 60%. Within one month after surgery， the patient experienced two sudden episodes of massive abdominal hemorrhage （>1 000 mL）， which were controlled by embolization of the right hepatic artery and adrenal artery， respectively. Subsequent treatments including cisplatin chemotherapy and disitamab vedotin combined with toripalimab were attempted， but liver metastasis still occurred after 7 weeks. The patient died 3 months after diagnosis. When elderly patients present with spontaneous renal rupture complicated by retroperitoneal hemorrhage， RA should be included in the differential diagnosis. Arterial embolization can be used for emergency hemostasis but is difficult to curb tumor progression， while current chemotherapy and immunotherapy regimens have limited efficacies， and there is an urgent need to explore targeted combination therapy.

Clear aligners have unique advantages in the treatment of Class Ⅱ malocclusion at mixed dentition stage， providing a more efficient and comfortable orthodontic option for clinical practice. The authors reported a clinical case of early treatment with clear aligners in a child patient at mixed dentition stage with Angle Class Ⅱ division 1 malocclusion accompanied by deep overbite and large overjet， and analyzed the clinical efficacy of this treatment method， so as to provide a reference for clinical diagnosis and treatment. The patient， a 7-year-old female， presented with parental chief complaint of “protruding teeth and prominent lips”. She was diagnosed as Angle Class Ⅱ division 1 malocclusion， convex facial profile， deep overbite， and large overjet. Imaging examination showed that the angle of long axis of maxillary central incisor to sella-nasion point line （U1-SN） was 114.46°， and the angle of subspinale-nasion-supramentale （ANB） was 5.39°. Clear aligners were used for treatment. After 17 months of treatment， the child patient achieved good dental arch form， tooth alignment and occlusion， facial profile， and soft tissue profile. The treatment effect was stable after 6 months of treatment. The application of clear aligners shows a good therapeutic effect in the child patient at mixed dentition stage with Angle Class Ⅱ division 1 malocclusion.

Objective To investigate the body weight monitoring management behaviors of residents in some areas of Changchun City in Jilin province， and to analyze the weight monitoring management behaviors and influencing factors， so as to provide the basis for enhancing residents’ awareness of weight monitoring and promoting self-management behaviors of weight. Methods The data were derived from the survey on chronic diseases and their risk factors in demonstration areas for chronic disease prevention and control in some areas of Changchun City from 2023 to 2024. A stratified random sampling method was used for sampling design； according to the administrative division and population distribution of the demonstration areas for chronic disease prevention and control in Changchun city， 6 representative districts or counties were selected； within each selected district or county， 48 communities were randomly selected； the permanent residents aged 18 years and above were investigated， with a total of 15 604 persons. Questionnaire survey was used to collect data on demographic characteristics， behavioral risk factors and prevalence of chronic diseases. Physical examination was performed using standard methods to measure indicators including height， body weight， waist circumference and blood pressure. SPSS 26.0 software was used for statistical analysis of the data. Categorical data were expressed as frequencies and percentages. χ² test was used for comparison between groups； multivariate Logistic regression analysis was used to analyze the influencing factors. Results The rate of weight measurement management of residents in some areas of Changchun City was 82.38% （12 855/15 064）； the percentage of measuring body weight once within one month on average was 32.36% （5 049/15 064）； the rate of not measuring body weight was 17.62% （2 748/15 064）. The multivariate analysis results showed that male， separated， widowed， and divorced persons were hindering factors for weight measurement （P<0.000 1）； higher education level， hypertension， hyperlipidemia， diabetes， diet mainly composed of vegetables and fruits， use of salt-restriction spoons， oil-restriction cups， and waist circumference tape， moderate-intensity physical activity for at least 30 minutes per day on 3 days per week， and the belief that overweight and obesity are related to chronic diseases were promoting factors for body weight measurement （P<0.000 1）. Conclusion Targeted health education activities should be continuously carried out to enhance the weight monitoring awareness and self-management ability of residents in this city.

Cell division cycle protein 6 （CDC6） is a member of the adenosine triphosphatase （ATPase） associated with diverse cellular activities（AAA+ATPase） family associated with a variety of cellular activities， which possesses ATPase activity and is highly conserved in eukaryotes. CDC6 is a key component of the pre-replication complex （pre-RC） and participates in the initiation of DNA replication. Meanwhile， it is also involved in regulating the S/M cycle checkpoints and modulating cell mitosis. CDC6 is abnormally expressed in a variety of malignant tumors and regarded as an oncogene. Its dysregulated expression leads to abnormal DNA replication and DNA damage， thereby affecting the occurrence and development of tumors. Moreover， a variety of drugs and regulatory factors have been confirmed to be able to inhibit the expression of CDC6 and block the growth of tumor cells. Therefore， CDC6 is a potential therapeutic target and prognostic biomarker. Based on the domestic and international research achievements of CDC6， this paper reviewed the structure and inhibitors of CDC6 protein， as well as its pathogenesis in glioma， prostate cancer（PCa）， breast cancer and many other tumors， so as to provide references for the treatment of malignant tumors.

Human immunodeficiency virus （HIV） is a kind of virus that attacks the host immune system and causes acquired immunodeficiency syndrome（AIDS） in infected individuals， which seriously endangers human health. Trans-activator of transcription （Tat）， a small non-structural protein encoded by HIV， binds to the trans-activation response element （TAR） of HIV to promote the transcription of viral genome， and this effect is closely related to multiple post-translational modifications of Tat protein. This paper reviewed the different types of post-translational modifications （phosphorylation， acetylation， ubiquitination and methylation） of HIV-1 Tat protein， and summarized the main action sites of related post-translational modifications on Tat protein as well as their regulatory mechanism on HIV-1 transcription and replication. Such modifications not only affect the structure and stability of Tat protein， but also regulate its transcriptional activity and its interaction with host proteins， and ultimately affect viral replication and latency. Therefore， in-depth study on post-translational modifications of Tat protein not only helps to understand the molecular mechanism of HIV infection， but also provides a theoretical basis for the development of novel antiviral therapeutic targets.

Berberine is a kind of small-molecule isoquinoline alkaloid extracted from Chinese herbal medicines such as Coptis chinensis， and its anti-tumor effect has attracted extensive attention. It exerts anti-tumor effects through multiple mechanisms， including arresting cell cycle， inducing tumor cell apoptosis， promoting autophagy， inhibiting tumor metastasis， and activating the anti-tumor immune response of the body. Berberine has the problems of low bioavailability and poor stability in vivo， which limit its clinical application. A variety of nanomaterials such as polymeric nanoparticles， metal-organic frameworks and liposomes have been innovatively applied to the delivery of berberine. This paper discussed the therapeutic effects and action mechanisms of berberine in various tumors， and analyzed the application status of berberine nano-drug delivery system in the field of cancer treatment， so as to provide theoretical basis and practical guidance for the further optimization and innovation of cancer treatment strategies， and promote the transformation of relevant research achievements into clinical application.

Low-dose radiotherapy （LDRT） generally refers to a radiotherapy modality with a single dose of 0.5-2.0 Gy or a total dose below 10.0 Gy. It reduces toxicity to normal tissues while retaining anti-tumor effects， and has unique biological effects such as immunomodulation， vascular normalization， and tumor microenvironment remodeling. However， tumor radioresistance severely restricts the clinical efficacy of LDRT. The development of tumor radioresistance involves multiple mechanisms， including overactivation of the DNA damage repair system， dysregulation of cell cycle checkpoints， activation of anti-apoptotic pathways， remodeling of the tumor microenvironment， imbalance of reactive oxygen species homeostasis， and epigenetic alterations. In recent years， various novel radiosensitization strategies have been developed to synergize with LDRT to inhibit radioresistance， including radiosensitization based on high-atomic-number nanomaterials， oxidative stress regulation targeting the glutathione system， metabolic reprogramming targeting key glycolytic enzymes， and immunomodulation combined with immune checkpoint inhibitors. These relevant strategies show particular application prospects in LDRT. This study elucidated the molecular mechanisms underlying tumor radioresistance， reviewed the applications and mechanisms of radiosensitization strategies， and analyzed the challenges in clinical translation from three dimensions： dose optimization， safety evaluation， and technological innovation， aiming to provide theoretical basis for the optimal design and clinical application of radiosensitization strategies.