Objective To discuss the effect of knock down of gene of kinesin family member 3B （KIF3B）， an important component of primary cilia （PC） of in mouse embryonic palatal mesenchymal on the autophagy level of cells （mEPMCs） cells， and to clarify its mechanism. Methods The mEPMCs from gestational day 14.5 C57BL/6J mice cultured in vitro were collected and divided into control group （administered normal saline）， empty lentivirus transfected cell group （sh-NC group） （administered lentivirus transfection）， KIF3B knockdown group （sh-KIF3B group） （administered KIF3B gene knockdown）， and KIF3B knockdown plus Smoothened receptor agonist （SAG） group （sh-KIF3B+SAG group） （administered KIF3B gene knockdown followed by SAG addition）， based on whether the KIF3B gene was knocked down and whether the SAG was used to activate the sonic hedgehog （Shh） signaling pathway and its downstream coreceptor Smo， with 5 rats in each group. Transmission electron microscope was used to observe the morphology and the number of autophagosomes/autolysosomes in the mEPMCs in various groups； Western blotting method was used to detect the expression levels of autophagy-related proteins Beclin-1 and p62， and the Shh signaling pathway proteins Shh and Smo in the mEPMCs in various groups. Results The transmission electron microscope observation results showed that compared with control group， the number of autophagosomes/autolysosomes in sh-KIF3B group was significantly increased （P<0.05）； compared with sh-KIF3B group， the number of autophagosomes/autolysosomes in the mEPMCs in sh-KIF3B+SAG group was significantly decreased （P<0.05）. The Western blotting results showed that compared with control group， the Beclin-1 protein expression level in the mEPMCs in sh-KIF3B group was significantly increased （P<0.05）， and the KIF3B， p62， Shh， and Smo protein expression levels were significantly decreased （P<0.01）； compared with sh-KIF3B group， the Shh， Smo， and p62 protein expression levels in the mEPMCs in sh-KIF3B+SAG group were significantly increased （P<0.01）， and the Beclin-1 protein expression level was significantly decreased （P<0.01）. Conclusion Knockdown of KIF3B gene can promote autophagy of the mEPMCs， and the mechanism may be related to its inhibition of the Shh signaling pathway.

Objective To discuss the role of angiopoietin-like protein 6 （ANGPTL6） in liver fibrosis， and to analyze the improving effect of engineered exosome（Exo）-delivered ANGPTL6 mRNA on liver fibrosis. Methods A total of 12 C57BL/6 mice were randomly divided into olive oil group （OIL group） （intraperitoneally injected with olive oil） and carbon tetrachloride （CCl₄） group （intraperitoneally injected with a mixture of olive oil and CCl?）， with 6 mice in each group； another 12 C57BL/6 mice were randomly divided into control group （fed a with methionine-choline sufficient diet） and methionine-choline deficient （MCD） group （fed a with MCD diet）， and two kinds of mouse liver fibrosis models were established. Real-time fluorescence quantitative PCR （RT-qPCR） and Western blotting method were used to detect the ANGPTL6 mRNA and protein expression levels in liver tissue of the mice in various groups. A total of 30 mice were randomly divided into olive oil + phosphate buffered saline （PBS） group （OIL+PBS group） （intraperitoneally injected with olive oil twice a week for 8 weeks， then injected with PBS buffer by tail vein twice a week for 6 weeks）， CCl₄+Exo-green fluorescent protein （GFP） mRNA group （established liver fibrosis model by intraperitoneal injection of CCl₄ mixture and were injected by tail vein with engineered Exo loaded with GFP mRNA for 6 weeks）， and CCl?+Exo-ANGPTL6 mRNA group （established liver fibrosis model by intraperitoneal injection of CCl₄ mixture and were injected by tail vein with engineered Exo loaded with ANGPTL6 mRNA for 6 weeks）， with 10 mice in each group. The mice in CCl₄+Exo-GFP mRNA group and CCl₄+Exo-ANGPTL6 mRNA group were injected with engineered Exo twice a week， 20 μg per mouse each time （volume 100 μL）. ELISA method was used to detect the serum alanine aminotransferase （ALT） and aspartate aminotransferase （AST） activities in the mice in various groups； Masson staining and Sirius red staining were used to observe the collagen deposition in liver tissue of the mice in various groups； immunohistochemistry method was used to detect the α-smooth muscle actin （α-SMA） expression levels in liver tissue of the mice in various groups； RT-qPCR method was used to detect the expression levels of α-SMA， collagen type Ⅰ alpha 1 chain （Col1a1）， transforming growth factor β1 （TGF-β1）， and tissue inhibitor of metalloproteinase 1 （TIMP-1） mRNA in liver tissue of the mice in various groups. Results The bioinformatics analysis results showed that ANGPTL6 expression was significantly down-regulated in activated hepatic stellate cell（aHSC）. The ultrasound examination results showed that the liver surface of the mice in OIL group was fine and smooth； compared with OIL group， the liver section of the mice in CCl? group was rough and uneven. The RT-qPCR and Western blotting results showed that compared with OIL group， the ANGPTL6 mRNA and protein expression levels in liver tissue of the mice in CCl? group were significantly decreased （P<0.05）. The engineered Exo extracted from the supernatant of HEK293T cells had intact structure and could be largely enriched in the fibrotic liver after tail vein injection， with GFP protein being largely expressed in the liver. The ELISA assay results showed that compared with OIL+PBS group， the ALT and AST activities in CCl₄+Exo-GFP mRNA group were significantly increased （P<0.05）； compared with CCl₄+Exo-ANGPTL6 mRNA group， the serum ALT and AST activities in CCl₄+Exo-GFP mRNA group were significantly decreased （P<0.05）. The Masson staining and Sirius red staining results showed that compared with OIL+PBS group， the collagen deposition in liver tissue of the mice in CCl?+Exo-GFP mRNA group was significantly increased， and the relative collagen area was increased （P<0.05）； compared with CCl₄+Exo-GFP mRNA group， the collagen deposition in tissue liver of the mice in CCl?+Exo-ANGPTL6 mRNA group was significantly decreased， and the relative collagen area was decreased （P<0.05）. The immunohistochemistry results showed that compared with OIL+PBS group， the α-SMA protein expression level in liver tissue of the mice in CCl?+ Exo-GFP mRNA group was significantly increased （P<0.05）； compared with CCl₄+Exo-GFP mRNA group， the α-SMA protein expression level in liver tissue of the mice in CCl?+Exo-ANGPTL6 mRNA group was significantly decreased （P<0.05）. The RT-qPCR results showed that compared with OIL+PBS group， the expression levels of Col1a1， α-SMA， TGF-β1， and TIMP-1 mRNA in liver tissue of the mice in CCl?+Exo-GFP mRNA group were significantly increased （P<0.05）； compared with CCl₄+Exo-GFP mRNA group， the expression levels of Col1a1， α-SMA， TGF-β1， and TIMP-1 mRNA in liver tissue of the mice in CCl?+Exo-ANGPTL6 mRNA group were significantly decreased （P<0.05）. Conclusion Engineered Exo-delivered ANGPTL6 mRNA injected via the tail vein in the mice is mainly enriched in the liver， and engineered Exo delivery of ANGPTL6 mRNA has an improving effect on liver fibrosis in the mice.

Objective To discuss the suitable concentration of D-galactose （D-gal） and modeling period， and establish its induced aging-related cognitive dysfunction model in the mice， and perform a comprehensive evaluation. Methods Fifty C57BL/6J mice were randomly divided into control group and 100， 200， 400， and 800 mg·kg^-1 D-gal groups， with 10 mice in each group. The mice in various D-gal groups were subcutaneously injected with the corresponding concentration of D-gal once daily； the mice in control group were injected with an equal volume of normal saline. The body mass and water consumption of the mice in various groups were monitored； forelimb grip strength test and experiment on the ability of pole climbing sports were used to evaluate the motor coordination ability of the mice in various groups； novel object recognition test， Y maze test， and Morris water maze test were used to evaluate the cognitive function of the mice in various groups； HE staining and Nissl staining were used to observe the pathomorphology of brain tissue of the mice in various groups； immunohistochemistry method was used to detect the expression of β-galactosidase （β-gal） protein in brain tissue of the mice in various groups； real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the mRNA expression levels of interleukin （IL）-1β， tumor necrosis factor-α （TNF-α）， IL-18， and IL-4 in hippocampus tissue of the mice in various groups； Western blotting method was used to detect the expression levels of β-gal， p53， and p16 proteins in hippocampus tissue of the mice in various groups. Results The body mass growth trends of the mice in control group and various D-gal groups were consistent and there was no statistically significant difference （P>0.05）， and there was no statistically significant difference in water consumption （P>0.05）. After 8 weeks of subcutaneous injection of D-gal， compared with control group， the forelimb grip strength values of the mice in 200 and 400 mg·kg?1 D-gal groups were significantly decreased （P<0.05 or P<0.01）； the pole-climbing time of the mice in 200 mg·kg?1 D-gal group was significantly prolonged （P<0.05）； the recognition indexes of the mice in 200 and 400 mg·kg?1 D-gal groups were significantly decreased （P<0.01）； the spontaneous alternation rate of the mice in 100， 200， 400， and 800 mg·kg?1 D-gal group was significantly decreased （P<0.05 or P<0.01）， the escape latency was significantly increased （P<0.05）.Spatial probe test showed that compared with control group， the escape latency of the mice in 200 mg·kg?1 D-gal group was significantly increased （P<0.05）. The HE staining and Nissl staining results showed that compared with control group， the hippocampus neurons of the mice in 200 mg·kg^-1 D-gal group were arranged disorderly， with obvious nuclear pyknosis， nuclear condensation， and abnormal morphology and structure， and the number of Nissl staining positive cells was significantly decreased. The immunohistochemistry results showed that compared with control group， the β-gal expressions in CA1 region， CA3 region， and cortex region of hippocampus tissue of the mice in 200 mg·kg?1 D-gal group were strongly positive. The RT-qPCR results showed that compared with control group， the expression levels of IL-1β，IL-18， and TNF-α mRNA in hippocampus tissue of the mice in 200 mg·kg?1 D-gal group were significantly increased （P<0.05 or P<0.01）， and the expression level of IL-4 mRNA was significantly decreased （P<0.01）. The Western blotting results showed that compared with control group， the expression levels of β-gal， p53， and p16 proteins in hippocampus tissue of the mice in 200 mg·kg?1 D-gal group were significantly increased （P<0.05 or P<0.01）. Conclusion The aging-related cognitive dysfunction model in the mice can be established by subcutaneous injection of 200 mg·kg?1 D-gal daily for 8 weeks.

Objective To discuss the effect of acupuncture on the differentiation and apoptosis of quadriceps muscle satellite cells in model rats with knee osteoarthritis（KOA）， and to clarify its related mechanism. Methods A total of 40 SPF-grade rats were selected and randomly divided into control group， model group， celecoxib group， and acupuncture group， with 10 rats in each group. The rats in control group only underwent joint cavity incision followed by suturing， while the rats in model group， celecoxib group， and acupuncture group were used to replicate the KOA models. The maximum circumference of the femoral segment of the affected limb， rat body mass， and quadriceps wet weight of the rats in various groups were measured； the quadriceps wet weight maintenance rate and quadriceps wet weight/body mass ratio of the rats in various groups were calculated. HE staining was used to observe the pathomorphology of articular cartilage and quadriceps muscle tissue of the rats in various groups； terminal deoxynucleotidyl transferase （TdT）-mediated dUTP nick end labeling （TUNEL） method was used to detect the apoptosis indexes in articular cartilage and quadriceps muscle tissue of the rats in various groups； immunofluorescence method was used to detect the protein expression levels of interleukin-6 （IL-6）， Janus kinase （JAK）， and signal transducer and activator of transcription 3 （STAT3） in quadriceps muscle tissue of the rats in various groups； Western blotting method was used to detect the expression levels of IL-6/JAK/STAT3 signaling pathway proteins， and muscle satellite cells， and apoptosis-related proteins in quadriceps muscle tissue of the rats in various groups. Results Compared with control group， the affected hind limb circumference， quadriceps wet weight， wet weight maintenance rate， and wet weight/body mass ratio of the rats in model group were significantly decreased （P<0.05）； compared with model group， the affected hind limb circumference， quadriceps wet weight， wet weight maintenance rate， and wet weight/body mass ratio of the rats in celecoxib group and acupuncture group were significantly increased （P<0.05）； compared with celecoxib group， the affected hind limb circumference， quadriceps wet weight， wet weight maintenance rate， and wet weight/body mass ratio of the rats in acupuncture group were significantly increased （P<0.05）. The HE staining results showed that the knee articular cartilage of the rats in control group remained intact， chondrocytes were aggregated and horizontally arranged with smooth edges， and quadriceps muscle cells were long cylindrical， orderly arranged， and regular in shape； in model group， the knee articular cartilage was thinner with rough edges， reduced number of cartilage layers， and disordered arrangement， and the quadriceps muscle fibers were disorganized， with some muscle fiber dissolution and muscle cell membrane damage， accompanied by muscle fiber fragments and a large amount of inflammatory exudate； in celecoxib group， the morphology of knee articular cartilage was generally normal， occasionally with irregular cartilage arrangement and reduced thickness， sporadically visible necrotic chondrocytes， quadriceps muscle fibers and sarcolemma were relatively intact， new muscle fibers appeared， some muscle fiber edges were blurred， accompanied by a small amount of cell debris and mild inflammatory infiltration； in acupuncture group， the knee articular cartilage structure remained intact with smooth edges， occasionally rough edges， and chondrocytes were aggregated and orderly arranged. The TUNEL assay results showed that compared with control group， the apoptosis indexes in articular cartilage and quadriceps muscle tissue of the rats in model group were significantly increased （P<0.05）； compared with model group， the apoptosis indexes in articular cartilage and quadriceps muscle tissue of the rats in celecoxib group and acupuncture group were significantly decreased （P<0.05）； compared with celecoxib group， the apoptosis index in articular cartilage and quadriceps muscle tissue of the rats in acupuncture group were significantly decreased （P<0.05）. The immunofluorescence assay results showed that compared with control group， the expression levels of IL-6， JAK， and STAT3 proteins in quadriceps muscle tissue of the rats in model group were significantly decreased （P<0.05）； compared with model group， the expression levels of IL-6， JAK， and STAT3 proteins in quadriceps muscle tissue of the rats in celecoxib group and acupuncture group were significantly increased （P<0.05）； compared with celecoxib group， the expression levels of IL-6， JAK， and STAT3 proteins in quadriceps muscle tissue of the rats in acupuncture group were significantly increased （P<0.05）. The Western blotting results showed that compared with control group， the expression levels of IL-6， JAK， STAT3， paired box transcription factor 7 （Pax7）， Desmin， Myosin， and Myogenin proteins in quadriceps muscle tissue of the rats in model group were significantly decreased （P<0.05）； compared with model group， the expression levels of IL-6， JAK， STAT3， Pax7， Desmin， Myosin， and Myogenin proteins in quadriceps muscle tissue of the rats in celecoxib group and acupuncture group were significantly increased （P<0.05）； compared with celecoxib group， the expression levels of IL-6， JAK， STAT3， Pax7， Desmin， Myosin， and Myogenin proteins in quadriceps muscle tissue of the rats in acupuncture group were significantly increased （P<0.05）. Compared with control group， the expression levels of B-cell lymphoma 2 （Bcl-2）， B-cell lymphoma-xl （Bcl-xl）， and myeloid cell leukemia 1 （MCL1） proteins in quadriceps muscle tissue in model group were significantly decreased （P<0.05）， and the expression levels of Bcl-2-associated X protein （Bax） and cysteinyl aspartate specific proteinase-3 （Caspase-3） proteins were significantly increased （P<0.05）； compared with model group， the expression levels of Bcl-2， Bcl-xl， and MCL1 proteins in quadriceps muscle tissue of the rats in celecoxib group and acupuncture group were significantly increased （P<0.05）， and the expression levels of Bax and Caspase-3 proteins were significantly decreased （P<0.05）； compared with celecoxib group， the expression levels of Bcl-2， Bcl-xl， and MCL1 proteins in quadriceps muscle tissue of the rats in acupuncture group were significantly increased （P<0.05）， and the expression levels of Bax and Caspase-3 proteins were significantly decreased （P<0.05）. Conclusion Acupuncture can promote the differentiation of quadriceps muscle satellite cells and inhibit muscle cell apoptosis in the model rats with KOA， and the mechanism may be related to the up-regulation of expressions of IL-6， JAK， and STAT3 proteins in the quadriceps muscle tissue.

Objective To study the effect of angiopoietin-1 （Ang-1） and tyrosine kinase receptor 2 （Tie2） inhibitor on glucose transportation in the human umbilical vein endothelial cells （HUVECs） cultured under high glucose conditions， and to clarify its mechanism. Methods The HUVECs were cultured in high glucose （30 mmol·L?1） in vitro and treated with 0， 200， 500， 1 000， and 2 000 μg·L?1 Ang-1 and 0， 2 500， 5 000， and 7 500 nmol·L?1 Tie2 inhibitor； cell counting kit-8 （CCK-8） method was used to detect the cell activity to screen the optimal concentrations of Ang-1 and Tie2 inhibitor. Glucose kit was used to detect the glucose level in the supernatant of the HUVECs after Ang-1 intervention. The HUVECs were randomly divided into blank control group （NG group）， high glucose group （HG group）， HG+Tie2 inhibitor group （HG+In-Tie2 group）， HG+Ang-1 group， HG+Ang-1+Tie2 inhibitor group （HG+Ang-1+In-Tie2 group）， and HG+Ang-1+phosphatidylinositol 3-kinase（PI3K） inhibitor group （HG+Ang-1+LY294002 group）. 5-Ethynyl-2'-deoxyuridine （EdU） method was used to detect the proliferation activities of the cells in various groups； YO-PRO-1/PI method was used to detect the apoptotic rates of the cells in various groups； real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of Ang-1 mRNA and Tie2 mRNA in the cells in various groups； Western blotting method was used to detect the expression levels of Tie2， glucose transporter 1 （GLUT1）， and glucose transporter 4 （GLUT4） proteins and the ratios of phosphorylated PI3K （p-PI3K）/PI3K and phosphorylated protein kinase B （p-AKT）/AKT in the cells in various groups. Results The CCK-8 assay results showed that compared with 0 μg·L?1 Ang-1 group， the activity of the HUVECs was significantly increased after treated with 200 μg·L?1 Ang-1 for 48 h （P<0.01）； compared with 0 nmol·L?1 Tie2 inhibitor group， the activity of the HUVECs was significantly decreased after treated with 2 500、 5 000 and 7 500 nmol·L?1 Tie2 inhibitor （P<0.01）； the optimal concentrations of Ang-1 and Tie2 inhibitor were 200 μg·L?1 and 2 500 nmol·L?1， respectively. Compared with NG group， the glucose level in the supernatant of the HUVECs in HG group was significantly increased （P<0.01）； compared with HG group， the glucose level in the supernatant of the HUVECs in Ang-1 group was significantly decreased （P<0.01）. The EdU assay results showed that compared with NG group， the proliferation activity of the HUVECs in HG group was significantly decreased （P<0.01）； compared with HG group， the proliferation activity of the HUVECs in HG+In-Tie2 group was significantly decreased （P<0.01）， and the proliferation activity of the HUVECs in HG+Ang-1 group was significantly increased （P<0.01）； compared with HG+Ang-1 group， the proliferation activities of the HUVECs in HG+Ang-1+In-Tie2 group and HG+Ang-1+LY294002 group were significantly decreased （P<0.01）. The YO-PRO-1/PI assay results showed that compared with NG group， the apoptotic rate of the HUVECs in HG group was significantly increased （P<0.01）； compared with HG group， the apoptotic rate of the HUVECs in HG+In-Tie2 group was significantly increased （P<0.01）， and the apoptotic rate of the HUVECs in HG+Ang-1 group was significantly decreased （P<0.01）； compared with HG+Ang-1 group， the apoptotic rates of the HUVECs in HG+Ang-1+In-Tie2 group and HG+Ang-1+LY294002 group were significantly increased （P<0.01）. The RT-qPCR results showed that compared with NG group， the expression levels of Ang-1 mRNA and Tie2 mRNA in the HUVECs in HG group and HG+In-Tie2 group were significantly decreased （P<0.01）； compared with HG group， the expression levels of Ang-1 mRNA and Tie2 mRNA in HG+ In-Tie2 group were significantly decreased （P<0.01）， and the expression levels of Ang-1 mRNA and Tie2 mRNA in the HUVECs in HG+Ang-1 group were significantly increased （P<0.05）； compared with HG+Ang-1 group， the expression levels of Ang-1 mRNA and Tie2 mRNA in the HUVECs in HG+Ang-1+In-Tie2 group and HG+Ang-1+LY294002 group were significantly decreased （P<0.05 or P<0.01）. The Western blotting results showed that compared with NG group， the expression level of Tie2 protein in the HUVECs in HG group was significantly decreased （P<0.01）， and the expression levels of GLUT1 and GLUT4 proteins were significantly increased （P<0.01）； compared with HG group， the expression levels of Tie2， GLUT1， and GLUT4 proteins in the HUVECs in HG+In-Tie2 group were significantly decreased （P<0.01）， the expression level of Tie2 protein in the HUVECs in HG+Ang-1 group was significantly increased （P<0.01）， and the expression levels of GLUT1 and GLUT4 proteins were significantly decreased （P<0.01）； compared with HG+Ang-1 group， the expression levels of Tie2， GLUT1， and GLUT4 proteins in the HUVECs in HG+Ang-1+In-Tie2 group and HG+Ang-1+LY294002 group were significantly decreased （P<0.01）. Compared with NG group， the p-PI3K/PI3K and p-AKT/AKT ratios in the HUVECs in HG group were significantly increased （P<0.01）； compared with HG group， the p-PI3K/PI3K and p-AKT/AKT ratios in the HUVECs in HG+In-Tie2 group were significantly decreased （P<0.01）， and the p-PI3K/PI3K and p-AKT/AKT ratios in the HUVECs in HG+Ang-1 group were significantly decreased （P<0.01）； compared with HG+Ang-1 group， the p-PI3K/PI3K and p-AKT/AKT ratios in the HUVECs in HG+Ang-1+In-Tie2 group and HG+Ang-1+LY294002 group were significantly decreased （P<0.01）. Conclusion Ang-1 down-regulates the expressions of GLUT1 and GLUT4 in the HUVECs cultured under high glucose conditions； the binding of Ang-1 to Tie2 may down-regulate GLUT1 and GLUT4 via the PI3K/AKT signaling pathway to participate in the glucose transportation in the HUVECs cultured under high glucose conditions.

Objective To discuss the improvement effect of epicatechin（EC） on acetaminophen（APAP）- induced liver injury in the mice， and to clarify its possible mechanism. Methods A total of 60 C57BL/6J mice were randomly divided into blank control group， APAP model group， low dose of EC group （10 mg·kg^-1）， middle dose of EC group （20 mg·kg^-1） and high dose of EC group （40 mg·kg?1）， with 12 mice in each group. Except for blank control group， the mice in the other groups were given intraperitoneal injection of APAP （200 mg·kg?1） to establish the liver injury models. At 1 h before APAP injection， the mice in low， middle and high doses of EC groups were intraperitoneally injected with 10， 20 and 40 mg·kg^-1 EC， respectively. A total of 36 nuclear factor E2-related factor 2 （Nrf2） deficient mice （Nrf2^-/- mice） were randomly divided into control group， APAP group and APAP+EC group， with 12 mice in each group. After modeling 24 h， the mice were sacrificed， and the blood and liver tissue of the mice were collected for subsequent detection. HE staining was used to observe the pathomorphology of the liver tissue in the mice in various groups； kit assay was used to detect the activities of aspartate aminotransferase （AST） and alanine aminotransferase （ALT） in serum of the mice in various groups and the myeloperoxidase （MPO） activity and the levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， malondialdehyde （MDA）， adenosine triphosphate （ATP）， glutathione （GSH） and ferrous ion （Fe2?） in liver tissue of the mice in various groups； Western blotting method was used to detect the expression levels of nuclear factor-κB （NF-κB） and Nrf2 signaling pathway-related proteins in liver tissue of the mice in various groups. Results The HE staining results showed that compared with APAP model group， the APAP-induced liver pathology injury in the mice in different doses of EC groups was significantly improved. Compared with blank control group， the levels of ALT and AST in serum of the mice in APAP model group were significantly increased （P<0.01）. Compared with APAP model group， the activities of ALT and AST in serum of the mice in low， middle and high doses of EC groups were significantly decreased （P<0.05 or P<0.01）. Compared with blank control group， the MPO activity and the levels of TNF-α and IL-1β in liver tissue of the mice in APAP model group were significantly increased （P<0.01）. Compared with APAP model group， the MPO activity and the levels of TNF-α and IL-1β in liver tissue of the mice in low， middle and high doses of EC groups were decreased （P<0.05 or P<0.01）. Compared with blank control group， the levels of MDA and Fe2? in liver tissue of the mice in APAP model group were significantly increased （P<0.05）， and the levels of ATP and GSH were significantly decreased （P<0.05）. Compared with APAP model group， the levels of MDA and Fe2? in liver tissue of the mice in low， middle and high doses of EC groups were significantly increased （P<0.05 or P<0.01）， and the levels of ATP and GSH were significantly increased （P<0.01）. Compared with blank control group， the expression levels of amino acid exchange transporter （xCT） and glutathione peroxidase 4 （GPX4） proteins in liver tissue of the mice in APAP model group were significantly decreased （P<0.05）； compared with APAP model group， the expression levels of xCT and GPX4 proteins in liver tissue of the mice in low， middle and high doses of EC groups were significantly increased （P<0.01）. Compared with blank control group， the expression levels of nuclear factor-κB （NF-κB） p-p65 and phosphorylated NF-κB inhibitor α （p-IκBα） proteins in liver tissue of the mice in APAP model group were significantly increased （P<0.05）； compared with APAP model group， the expression levels of NF-κB p-p65 and p-IκBα proteins in liver tissue of the mice in low， middle and high doses of EC groups were significantly decreased （P<0.01）. Compared with blank control group， the expression levels of Nrf2 and heme oxygenase-1 （HO-1） proteins in liver tissue of the mice in APAP model group were significantly increased （P<0.05）； compared with APAP model group， the expression levels of Nrf2 and HO-1 proteins in liver tissue of the mice in low， middle and high doses of EC groups were significantly increased （P<0.01）. Compared with control group， the ALT level in serum and the levels of MDA and Fe²⁺ in liver tissue of the Nrf2^-/- mice in APAP group were significantly increased （P<0.01）， and the levels of ATP and GSH in liver tissue were significantly decreased （P<0.01）. Conclusion EC can improve APAP-induced liver injury in the mice， and its mechanism may be related to the inhibition of ferroptosis by activating the Nrf2/GPX4 signaling pathway.

Objective To explore the effects of imperatorin （IMP） on airway remodeling in the bronchial asthma mice， and to elucidate the possible mechanisms. Methods Forty SFP male BALB/c mice were randomly divided into control group， model group， low dose of IMP group （IMP-L group）， high dose of IMP group （IMP-H group） and dexamethasone group， with 8 mice in each group. Except for contol group， the mice in the other groups were injected with an ovalbumin （OVA） suspension intraperitoneally to induce the asthma models. After one week， the daily asthma symptoms of the mice were observed and scored. After 8 weeks， the enhanced pause（Penh） values of the mice in various groups were detected to evaluate the airway reactivities. The percentages of eosinophils in the bronchoalveolar lavage fluid （BALF） of the mice in various groups were detected by flow cytometry. The levels of serum IgE， interleukin interferon-gamma（IL）-13， IL-5， IL-4 and interferon-gamma（IFN-γ） in BALF of the mice in various groups were measured by enzyme-linked immunosorbent assay （ELISA） method. HE， PAS and Masson staining were applied to observe the pathomorphology， the number of goblet cells and collagen deposition of the lung tissue of the mice in various groups.Immunohisto chemistry method was applied to detect the expressions of α-smooth muscle actin （α-SMA） and mouse mammary tumor virus（MMTV） wingless type MMTV intergration site family member 5A （Wnt5A） proteins in lung tissue of the mice in various groups. The expression levels of Wnt5A， cellular myelocytomatosis oncogene （c-Myc）， β-catenin and α-SMA in lung tissue of the mice in various groups were detected by Western blotting method. The expression levels of α-SMA protein in lung tissue of the mice in various groups were detected by immunofluorescence method. Results Compared with control group， the score of asthma symptoms of the mice in model group was increased （P<0.01）； the Penh value was significantly increased （P<0.01）； the serum IgE levels and the levels of IL-13， IL-5， IL-4 in BALF， as well as the percentage of eosinophils （EOS） in BALF were significantly increased （P<0.05 or P<0.01）， and the level of IFN-γ was reduced （P<0.05）； the expression levels of α-SMA and Wnt5A proteins in lung tissue were markedly increased （P<0.01）； the expression levels of proteins associated with the Wnt/β-catenin signaling pathway in the lung tissue were significantly increased （P<0.01）； the immofluorescence method results showed the expression level of α-SMA protein in lung tissue was significantly increased （P<0.01）. Compared with model group， the scores of asthma symphtoms of the mice in IMP-L group， IMP-H group， and dexamethasone group were decereased （P<0.01）， and the Penh values of the mice in IMP-H group were decreased （P<0.05）； the serum IgE levels and the levels of IL-13， IL-5， IL-4 in BALF， as well as the percentages of EOS in BALF of the mice in IMP-L group， IMP-H group， and dexamethasone group were decreased （P<0.05 or P<0.01）， and the levels of IFN-γ were increased （P<0.05）； the expression levels α-SMA and Wnt5A proteins in lung tissue were decreased （P<0.05 or P<0.01）； the expression levels of proteins related to the Wnt/β-catenin signaling pathway in the lung tissue were decreased （P<0.05 or P<0.01）； the immunofluorescence method results showed that expression levels of the α-SMA protein in the lung tissue were reduced （P<0.05 or P<0.01）. Conclusion IMP has an improving effect on airway remodeling in the asthmatic mice and can inhibit the expression levels of Wnt/β-catenin pathway-related proteins.

Objective To investigate the effects of cholinergic receptor nicotinic alpha 5 subunit（CHRNA5） on the invasion and proliferation of pancreatic ductal adenocarcinoma （PDAC） using bioinformatics approaches and cellular experiments， and to analyze its regulatory pathways and provide foundation for the identification of novel therapeutic targets for pancreatic cancer. Methods Transcriptome， gene variation， and clinical data of pan-tumor and normal tissues were downloaded from public databases including The Cancer Genome Atlas （TCGA） and Genotype-Tissue Expression （GTEx）. The expression levels of CHRNA5 mRNA between pan-tumor patients and healthy individuals were compared using Wilcoxon test. The expression levels of CHRNA5 mRNA between PDAC and para-cancer tissues were further compared using datasets from the Gene Expression Omnibus （GEO） database. The data were divided into three groups according to the different expression levels of CHRNA5 mRNA using tertile method： CHRNA5 high expression group， CHRNA5 medium expression group， and CHRNA5 low expression group. Survival analysis was performed in CHRNA5 high expression and low expression groups to assess the impact of CHRNA5 in prognosis of pancreas cancer. Real-time fluorescence quantitative PCR （RT-qPCR） and immunohistochemistry methods were used to detect the expressions of CHRNA5 mRNA and protein in tumor tissue， paracancer tissue， tumor cells and normal cells. Differential analysis of the transcriptome data and enrichment analysis of Hallmark genes and Reactome pathways were then conducted. Immune cell infiltration analysis was performed on PDAC based on transcriptome data. Tumor mutation burden （TMB） analysis， single nucleotide variation （SNV）， and copy number variation （CNV） analysis were also conducted. The small interfering RNA （si）-NC and si-CHRNA5 were transfected into the MIA PaCa-2 and Capan-2 cells， and the samples were divided into NC， siRNA-1 and siRNA-2 groups according to the transfected siRNA. The proliferation activities of the cells in various groups under acetylcholine （Ach） and non-ACh conditions were detected using cell counting kit-8 （CCK-8） method， and the expression of c-Myc protein in the cells in various groups were detected using Western blotting method. Results Compared with normal tissue， the expression levels of CHRNA5 mRNA were elevated in 28 types of tumor tissues， including pancreatic cancer （adjusted P<0.05）. The results of RT-qPCR and immunohistochemistry revealed that CHRNA5 was highly expressed in tumor tissue and pancreatic ductal adenocarcinoma （PDAC） cells compared with paracancer tissue and normal pancreatic ductal cells. The survival analysis showed that the PDAC patients with high expression of CHRNA5 had worse prognosis compared with those with low expression of CHRNA5. A total of 994 differentially expressed genes （DEGs） were identified between PDAC and control samples， with 381 up-regulated genes and 613 down-regulated genes. The gene Set Enrichment Analysis （GSEA） results indicated that CHRNA5 was highly associated with oxidative phosphorylation， cell proliferation， immune infiltration， and cell cycle. The immune analysis results showed that the samples with high expression of CHRNA5 had poorer immune infiltration compared with those with low expression of CHRNA5. Compared with CHRNA5 low expression group， the mutation rates of pancreatic cancer progression-related genes KRAS， TP53， and SMAD4 in CHRNA5 high expression group were increased. In cell experiments， compared with NC+Ach group， the proliferation activities of the cells in siRNA-1+Ach group and siRNA-2+Ach group were significantly decreased （P<0.05）. The results of Western blotting revealed that the expression amounts of c-Myc protein in siRNA-1 and siRNA-2 groups were significantly lower than those in NC group. Conclusion CHRNA5 modulates the cell cycle via myelocytomatosis viral oncogene （MYC） family， thereby influencing the proliferation of pancreas cancer cells and promoting the progression of PDAC， suggesting that CHRNA5 is a potential therapeutic target for pancreas cancer.

Objective To explore the effect of heme-binding protein 1（HEBP1） down-regulation on the function of microglia BV2， and to clarify the key role of HEBP1 in the microglia. Methods Negative control and HEBP1 knockdown small interfering RNA （siRNA） were constructed to knockdown HEBP1 gene in mouse-derived microglial BV2， and the HEBP1 knockdown BV2 cell models were obtained. The BV2 cells were divided into si-NC group， si-HEBP1-1 group， si-HEBP1-2 group， and si-HEBP1-3 group. Real-time fluorescence quantitative PCR（RT-qPCR） and Western blotting methods were used to detect the expression levels of HEBP1 mRNA and protein in the BV2 cells after knockdown； the siRNA with the most significart knockdown effect was selected for stlbsequent expreriments.The proliferation abilities of the cells in si-NC group and si-HEBP1 group were detected by cell counting kit-8（CCK-8） assay， and the cell migration rates were assessed by scratch assay； the cellular mitochondrial membrane potential and reactive oxygen species （ROS） levels were detected by kits； the cellular mitochondrial respiratory function was detected by mitochondrial respirometer. The BV2 cells were divided into si-NC group， si-NC+lipopolysacch aride（LPS） group， si-HEBP1 group， and si-HEBP1+LPS group. RT-qPCR method was used to detect the expression levels of HEBP1， interleukin-1β（IL-1β），tumor necrosis factor-α（TNF-α），and interleukin-6（IL-6） mRNA in the BV2 cells in various groups， and Western blotting method was used to detect the expression levels of HEBP1 protein in the BV2 cells in various groups. Results When the BV2 cells were transfected with siRNA carrying with red fluorescence tag CY3， the transfect effricacy was above 90%； compared with si-NC group， the expression levels of HEBP1 protein in the BV2 cells in si-HEBP1-1 group， si-HEBP1-2 group， and si-HEBP1-3 group were significantly decreased （P<0.05 or P<0.01）， especially in si-HEBP1-1 group. Compared with si-NC group， the expression levels of HEBP1 mRNA in the BV2 cells in si-HEBP1-1 group， si-HEBP1-2 group， and si-HEBP1-3 group were significantly decreased （P<0.01）， especially in si-HEBP1-1 group； indicating that si-HEBP1-1 was the siRNA with best HEBP1 knowdown effect， and the HEBP1 knockdown BV2 cell model was successfully constructed. The CCK-8 resuts showed that compared with si-NC group， the proliferation activities of the BV2 cells in si-HEBP1 group were decreased （P<0.05 or P<0.01）； from 90 min， the differences in proliferation activities of the BV2 cells in two groups were obvious. The cell scratch experiment results showed that compared with si-NC group， the cell migration rate in si-HEBP1 group was significantly decreased （P<0.05）. The fluorescence microscope results showed that compared with si-NC group， the mitochondrial membrane potential of the BV2 cells in si-HEBP1 group was significantly decreased （P<0.05）； compared with si-NC group， the ROS level in the BV2 cells in si-HEBP1 group was significantly increased（P<0.05）. The mitochondrial respiration function testing results showed that compared with si-NC group， routine respiration（ROUNTINE） and leak respiration（LEAK） in si-HEBP1 group were significautly decreased （P<0.05 or P<0.01）， and electron transfer system capacity（ETS） and residual oxygen consumption（ROX） had no significant differences （P>0.05）； the ATP amount was decreased （P<0.05）. The RT-qPCR results showed that compared with si-NC group， the expression levels of IL-1β， TNF-α， and IL-6 mRNA in the BV2 cells in si-NC+LPS group were significantly decreased （P<0.01）； compared with si-HEBP1 group， the expression levels of IL-1β， TNF-α， and IL-6 mRNA in the BV2 cells in si-HEBP1+LPS group were significantly decreased （P<0.01）； compared with si-NC+LPS group， the expression levels of IL-1β， TNF-α， and IL-6 mRNA in the BV2 cells in si-HEBP1+LPS group were significantly increased （P<0.01）. Conclusion Knockdown of HEBP1 gene can decrease the proliferation and migration abilities of the microglia BV2 and increase inflammatory response to LPS stimulus， and their mechanisms may be related to mitochondrial function damage and decreased ATP production of the BV2 cells.

Objective To observe the effects of adipose-derived stem cells （ADSC） combined with acellular scaffold （AS） on the ultrastructure of dorsal root ganglion and the protein and mRNA expression levels of ciliary neurotrophic factor （CNTF）， Janus kinase 2 （JAK2）， phosphorylated JAK2 （p-JAK2）， signal transducer and activator of transcription 3（STAT3） and phosphorylated STAT3（p-STAT3） in the rats with sciatic nerve injury （SNI）， and to clarify the protective effect of ADSC combined with AS on dorsal root ganglion in the SNI rats and its possible mechanism. Methods The rat ADSCs were isolated and cultured and their multidirectional differentiation potential was detected. The AS of rats was prepared， and ADSCs were injected into the AS to construct tissue-engineered nerve. A total of 36 rats were randomly divided into control group， model group， AS group， and ADSC+AS group. The rats in control group were routinely fed， and the rats in other groups were used to establish the SNI models by resecting 10 mm of right sciatic nerve. The rats in model group received no further treatment， while the rats in AS group and ADSC+AS group were bridged with AS and the constructed tissue-engineered nerve at the two ends of the injured nerve， respectively. At 6 weeks after surgery， transmission electron microscope was used to observe the ultrastructure of dorsal root ganglion of the rats in various groups； immunofluorescence method was used to detect the protein expression levels of CNTF， p-JAK2， and p-STAT3 in dorsal root ganglion of the rats； real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the mRNA expression levels of CNTF， JAK2， and STAT3 in dorsal root ganglion of the rats in various groups. Results After 7 d of primary ADSC culture， a large number of large and long spindle-shaped cells were observed under the inverted microscope， arranged in clusters or whirlpools； red lipid droplets were observed with oil red O staining under microscope， and calcified nodules were observed with Alizarin red staining under microscope， indicating that the isolated and cultured cells had multidirectional differentiation ability.Compared with normal nerve tissue， the level of DNA in AS of rats was significantly decreased （P<0.05）. Compared with control group， the nuclear membrane of dorsal root ganglion cells in model group was uneven and serrated， the number of organelles in the cytoplasm was decreased， mitochondria were swollen with broken or missing cristae and unclear structure； the CNTF protein and mRNA expression levels were significantly decreased （P<0.01）， the p-JAK2 and p-STAT3 protein expression levels were significantly increased （P<0.01）， and the JAK2 and STAT3 mRNA expression levels were significantly increased （P<0.01）. Compared with model group， the serrated change of nuclear membrane of the dorsal root ganglion cells in AS group was significantly alleviated， the number of organelles in the cytoplasm was increased， and mitochondrial swelling was reduced； in ADSC+AS group， the nuclear membrane of dorsal root ganglion cells tended to be intact， the number of organelles was increased， and mitochondrial swelling and vacuolization were significantly reduced； the CNTF protein and mRNA expression levels in the dorsal root ganglion in AS group and ADSC+AS group were significantly increased （P<0.01）， the p-JAK2 and p-STAT3 protein expression levels were significantly decreased （P<0.01）， and the JAK2 and STAT3 mRNA expression levels were significantly decreased （P<0.01）. Compared with AS group， the CNTF protein and mRNA expression levels in ADSC+AS group were significantly increased （P<0.05 or P<0.01）， the p-JAK2 and p-STAT3 protein expression levels were significantly decreased （P<0.01）， and the JAK2 and STAT3 mRNA expression levels were significantly decreased （P<0.01）. Conclusion The application of ADSC combined with AS can improve the ultrastructure of dorsal root ganglion in the SNI rats， and the mechanism may be related to the increased CNTF expression and decreased activation of the JAK2/STAT3 signaling pathway in the dorsal root ganglion by ADSC combined with AS application.

Objective To discuss the effect of Compound Rehmannia Granules on intestinal flora of the pneumonia model rats induced by Streptococcus pneumoniae （Spn） through 16s rRNA sequencing technology， and to clarify its potential mechanism. Methods A total of 30 rats were randomly divided into control group （normal rats）， model group （Spn-induced pneumonia rat models）， low dose of Compound Rehmannia Granules group （pneumonia rat models administered 1.75 g·kg^-1 Compound Rehmannia Granules by gavage）， medium dose of Compound Rehmannia Granules group （pneumonia rat models administered 3.50 g·kg^-1 Compound Rehmannia Granules by gavage）， and high dose of Compound Rehmannia Granules group （pneumonia rat model administered 7.00 g·kg^-1 Compound Rehmannia Granules by gavage）， with 6 rats in each group. The wet/dry weight （W/D） ratio of lung tissue and blood gas indexes were measured； HE staining was used to observe the pathomorphology and the degree of lung injury in lung tissue of the rats in various groups were assessed； kit assay was used to detect the bacterial load level and the levels of interleukin （IL）-6， IL-8， and IL-10 in bronchoalveolar lavage fluid （BALF） of the rats in various groups； 16s rRNA intestinal flora sequencing analysis was performed. Results Compared with control group， the arterial partial pressure of carbon dioxide （PaCO₂） of the rats in model group was significantly increased （P<0.05）， and the arterial partial pressure of oxygen （PaO₂） and oxygen saturation （SaO₂） were significantly decreased （P<0.05）； compared with model group， the PaCO₂ of the rats in low， medium， and high doses of Compound Rehmannia Granules groups was significantly decreased （P<0.05）， and the PaO? and SaO? were significantly increased （P<0.05）， showing a dose-dependent manner. The HE staining results showed no significant injury in lung tissue of the rats in control group； the cells in lung tissue of the rats in model group were arranged disorderly with massive inflammatory cell infiltration， and the alveolar wall capillaries were significantly dilated； compared with model group， the morphological damage of lung tissue of the rats in low， medium， and high doses of Compound Rehmannia Granules groups was improved. Compared with control group， the W/D value and pathological score of lung tissue of the rats in model group were significantly increased （P<0.05）； compared with model group， the W/D value and pathological scores of lung tissue of the rats in low， medium， and high doses of Compound Rehmannia Granules groups were significantly decreased （P<0.05）， showing a dose-dependent manner. Compared with control group， the bacterial load level in BALF of the rats in model group was significantly increased （P<0.05）； compared with model group， the bacterial load level in BALF of the rats in low， medium， and high doses of Compound Rehmannia Granules groups was significantly decreased （P<0.05）， showing a dose-dependent manner. Compared with control group， the levels of IL-6 and IL-8 in BALF of the rats in model group were significantly increased （P<0.05）， and the IL-10 level was significantly decreased （P<0.05）； compared with model group， the levels of IL-6 and IL-8 in BALF of the rats in low， medium， and high doses of Compound Rehmannia Granules groups were significantly decreased （P<0.05）， and the IL-10 level was significantly increased （P<0.05）， showing a dose-dependent manner. Compared with control group， the microbial abundance indicator（Chao1）， microbial diversity and evenness indicator（Shannon）， microbial dominance indicator （Simpson）， and observed species indicator （observed_species） of the rats in model group were significantly decreased （P<0.05）； compared with model group， the Chao1， Shannon， Simpson， and （observed_species） indices of the rats in low， medium， and high doses of Compound Rehmannia Granules groups were significantly increased （P<0.05）， showing a dose-dependent manner. Compared with control group， the relative abundance of Bacteroidetes in model group was decreased， the relative abundance of Firmicutes was increased， and the Firmicutes/Bacteroidetes ratio was significantly increased （P<0.05）； compared with model group， the relative abundances of Bacteroidetes in low， medium， and high doses of Compound Rehmannia Granules groups were increased， the relative abundance of Firmicutes was decreased， and the Firmicutes/Bacteroidetes ratio was significantly decreased （P<0.05）， showing a dose-dependent manner. At the family level， compared with control group， the relative abundances of Corynebacteriaceae， Staphylococcaceae， and Moraxellaceae in model group were significantly increased， while the relative abundances of Lactobacillaceae， Lachnospiraceae， and Akkermansiaceae were significantly decreased； compared with model group， the relative abundances of Corynebacteriaceae， Staphylococcaceae， and Moraxellaceae in low， medium， and high doses of Compound Rehmannia Granules groups were significantly decreased， and the relative abundances of Lactobacillaceae， Lachnospiraceae， and Akkermansiaceae were significantly increased. At the genus level， compared with control group， the relative abundances of Desulfovibrio and Facklamia in model group were significantly increased， and the relative abundances of Bifidobacterium and Ruminococcaceae were significantly decreased； compared with model group， the relative abundances of Desulfovibrio and Facklamia in low， medium， and high doses of Compound Rehmannia Granules groups were significantly decreased， and the relative abundances of Bifidobacterium and Ruminococcaceae were significantly increased. Conclusion Compound Rehmannia Granules can alleviate inflammation and lung injury in Spn-induced pneumonia rats， which may be related to the increase in the abundance and diversity of intestinal flora.

Objective To discuss the role of circular RNAs （circRNAs）-overexpressing modified dental pulp stem cells （DPSCs）-derived exosomes （Exo） in promoting angiogenesis of the human umbilical vein endothelial cells （HUVECs） in dental pulp vascular regeneration， and to clarify the related molecular mechanism. Methods The primary DPSCs derived from deciduous teeth， adult wisdom teeth， and elderly permanent teeth were isolated. Flow cytometry was used to detect the positive expression of surface marker proteins in three kinds of primary DPSCs. Co-culture systems were established using three sources of DPSCs and HUVECs， and the cells were divided into deciduous teeth-derived primary DPSCs group， adult wisdom teeth-derived primary DPSCs group， and elderly permanent teeth-derived primary DPSCs group. Additionally， the cells were divided into Exo-OE vector group， Exo-circ_0026827 OE group， and Exo-circRNA124534 OE group； after transfection with adenovirus-mediated OE vector， circ_0026827 OE， and circRNA124534 OE， respectively， the Exo from the cell culture supernatant were isolated. Cell counting kit-8 （CCK-8） method was used to detect the proliferation activities of the HUVECs in various groups； real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of angiogenesis-related circRNAs in the Exo in various groups； Western blotting method was used to detect the expression levels of Exo marker-related proteins in the supernatant of co-cultured cells. The uptakes of Exo by the cells in various groups were verified； angiogenesis induction experiment was used to detect the isolation of Exo from cell supernatant. Results Most cells expressed CD44（+）， CD34（-）， and Stro-1（+）， and were identified as primary DPSCs.The CCK-8 assay results showed that compared with deciduous teeth-derived primary DPSCs group， the proliferation activity of the HUVECs co-cultured in adult wisdom teeth-derived primary DPSCs group and elderly permanent teeth-derived primary DPSCs group was significantly decreased （P<0.01）； compared with adult wisdom teeth-derived primary DPSCs group， the proliferation activity of the HUVECs co-cultured in elderly permanent teeth-derived primary DPSCs group was significantly decreased （P<0.01）. cluster of differentiation 9 （CD9）， heat shock protein 70 （HSP70）， and tumor susceptibility gene 101（TSG101） were positively expressed in the Exo from the cell culture supernatant of three co-culture systems. The particle sizes of Exo from the cell supernatant in three co-culture systems were 50-110 nm. The RT-qPCR results showed that compared with deciduous teeth-derived primary DPSCs group， the mRNA expression levels of circRNA124534 and circ_0026827 in the Exo in adult wisdom teeth-derived primary DPSCs group and elderly permanent teeth-derived primary DPSCs group were significantly decreased （P<0.01）； compared with adult wisdom teeth-derived primary DPSCs group， the mRNA expression levels of circRNA124534 and circ_0026827 in the Exo in elderly permanent teeth-derived primary DPSCs group were significantly decreased （P<0.01）； there was no statistically significant difference in the circ_signal-induced proliferation-associated 1 like 1 （SIPA1L1） mRNA expression level among three groups （P>0.05）. The Western blotting results showed that compared with Exo-OE vector group， the protein expression levels of phosphorylated p38 mitogen-activated protein kinase （p-p38 MAPK）， vascular endothelial growth factor A （VEGF-A）， vascular endothelial growth factor receptor 2 （VEGFR2）， angiopoietin 1 （Ang-1）， stromal cell-derived factor 1 （SDF-1）， and matrix metalloproteinase 9 （MMP-9） in the HUVECs in overexpression circ_0026827 and circRNA124534 were significantly increased （P<0.01）； compared with Exo-OE vector group， the protein expression levels of p-p38 MAPK， VEGF-A， VEGFR2， Ang-1， SDF-1， and MMP-9 in the HUVECs in Exo-circRNA124534 OE group were significantly increased （P<0.01）. The HUVECs in various groups successfully took up Exo.The HUVECs in Exo-OE vector group showed a relatively flattened growth state. Compared with Exo-OE vector group， the HUVECs in Exo-circ_0026827 OE group and Exo-circRNA124534 OE group showed a reticularly arranged growth state and a tendency to form tubular structures. Conclusion The deciduous teeth DPSCs-derived Exo modified by overexpression circ_0026827 and circRNA124534 has a certain promoting effect on angiogenesis of the HUVECs.

Objective To discuss the optimal doping concentration of vanadium （V） and silicon （Si） co-doped TiO? coating （V-Si TiO?） formed on titanium surface by electrochemical treatment， to evaluate its antibacterial effect under visible light irradiation， and to clarify its visible light response mechanism. Methods The medical pure titanium sheets were subjected to micro-arc oxidation followed by high-temperature calcination， and V-Si TiO₂ coatings with different doping concentrations were prepared by adjusting the ratio of V to Si in the electrolyte. The experiment was divided into 1V∶10Si （V5Si50） group， 2V∶10Si （V10Si50） group， and 3V∶10Si （V15Si50） group； control group was set up（contains only bacterial culture medium）. The optimal doping concentration was screened based on comprehensive evaluation of surface morphology， ion release， photocatalytic ability， and biocompatibility； cell counting kit-8（CCK-8） method was used to detect the proliferation activities and the survival rates of the cells in various group. Subsequently， the optimized coating was characterized and compared by scanning electron microscope （SEM）， atomic force microscopy （AFM）， digital eddy current coating thickness gauge， X-ray diffraction （XRD）， X-ray photoelectron spectroscope （XPS）， and ultraviolet-visible absorption spectroscopy （UV-vis）. The experiment was divided into PT group （blank control）， PEO group （no element doping）， V10 group （V doping）， Si50 group （Si doping）， and V10Si50 group （2V∶10Si）. The ability of the coating materials to degrade methylene blue （MB） and generation of reactive oxygen species （ROS） under visible light were detected. For antibacterial experiments， Staphylococcus aureus （S.aureus） and Escherichia coli （E.coli） were used. The colony counts on plates in various groups were recorded after visible light irradiation for 2 h and dark treatment for 2 h， respectively. The ROS levels were detected using 2'，7'-dichlorofluorescein diacetate （DCFH-DA） ROS probe. ROS scavenging experiment was performed using the optimal doping concentration V10Si50 group， and the two kinds of bacteria were divided into blank control group， N-acetylcysteine （NAC） group， V10Si50 group， and NAC+V10Si50 group. The colony counts on plates in various groups were recorded after visible light irradiation for 2 h. Results The V concentration of 0.01 mol·L?1 and Si concentration of 0.05 mol·L?1 in the electrolyte solution were the optimal doping concentrations for the V-Si TiO? coating. The SEM observation results showed that compared with V5Si50 group and V15Si50 group， the surface pore size of the coating material in V10Si50 group was significantly decreased （P<0.05）， and the coating thickness was significantly increased （P<0.05）； its crystal structure was mainly anatase type， and the MB degradation rate of the coating material in V10Si50 group after 9 h of visible light catalysis was significantly increased （P<0.05）. Compared with control group， the cell proliferation activity and cell survival rate in V10Si50 group were significantly increased at 1， 2， and 4 d of cell culture （P<0.05）； at 2 and 4 d of cell culture， the cell proliferation activity and cell survival rate in V5Si50 group and V15Si50 group were significantly decreased （P<0.05）. Compared with PT， PEO， and Si50 groups， the colony counts of two kinds of the bacteria in V10 group and V10Si50 group after visible light irradiation for 2 h were significantly decreased （P<0.05）. Compared with PT group and PEO group， the ROS levels in two kinds of the bacteria in V10Si50 group after 2 h of irradiation were significantly increased （P<0.05）. Compared with V10Si50 group， the colony counts of two kinds of the bacteria in NAC+V10Si50 group were significantly increased （P<0.05）. Conclusion A reasonably loaded V-Si TiO? coating material （V10Si50） was screened out， which maintained good biological activity and significantly enhanced the antibacterial effect under visible light irradiation.

Objective To discuss the changes of early memory T lymphocytes （T_M） in the mice infected with influenza A virus （H1N1） PR8， and to clarify its significance. Methods A total of 32 C57BL/6 mice were randomly divided into control group and influenza virus （PR8） group， with 16 mice in each group. The mice in control group were intranasally administered 30 μL of sterile phosphate buffered saline （PBS）， and the mice in PR8 group were intranasally administered 2 LD₅₀ PR8 virus solution. The mice were sacrificed on the 10 th day， and bronchoalveolar lavage fluid （BALF）， lung tissue， and lymphnodes were collected for subsequent experiments. HE staining was used to observe the pathomorphology of lung tissue in the mice in two groups； hemagglutination assay， real-time fluorescence quantitative PCR （RT-qPCR） method， and immunofluorescence method were used to detect the viral infection in lung tissue of the mice； flow cytometry was used to detect the percentages of T lymphocytes， effector T lymphocytes， central memory T lymphocytes （T_CM）， effector memory T lymphocytes （T_EM）， and nuclear protein（NP）/pdymerase acidic protein（PA）-specific T_EM lymphocytes in BALF， lung tissue， and lymphnodes of the mice in two groups. Results Compared with control group， the lung tissue of the mice in PR8 group showed significant pathological damage， and the lung tissue virus titer and NP protein expression level were significantly increased （P<0.01）. Compared with control group， the percentages of CD4⁺ T lymphocytes， CD8⁺ T lymphocytes， effector CD4⁺ T lymphocytes， effector CD8⁺ T lymphocytes， CD4⁺ T_EM lymphocytes， and CD8⁺ T_EM lymphocytes in BALF and lung tissue of the mice in PR8 group were significantly increased （P<0.01）. Compared with control group， the positive percentages of CD4⁺ T_CM， CD8⁺ T_CM， CD4⁺ T_EM， CD8⁺ T_EM， and NP/PA-specific CD4⁺ T_EM， CD8⁺ T_EM lymphocytes in BALF and lung tissue of the mice in PR8 group were significantly increased （P<0.05 or P<0.01）， and the percentage of CD8+ T_CM lymphocytes in the lymph nodes was significantly increased （P<0.05）. Conclusion Influenza virus infection can induce the proliferation and differentiation of T lymphocytes into effector T lymphocytes， leading to the early establishment of virus-specific T_M lymphocytes targeting the conserved NP/PA epitopes in the respiratory mucosa.

Objective To express， purify， prepare antibodies， and analyze the subcellular localization of Toxoplasma gondii thioredoxin 20（Trx20）， and to provide the reference for the development of Toxoplasma gondii vaccine. Methods Bioinformatics-related websites and software were used to perform bioinformatics analysis of the Trx20 protein； specific primers were designed to amplify the target fragment and construct the prokaryotic expression vector； the protein was expressed in vitro and purified； experimental animals were immunized to prepare antibodies； enzyme-linked immunosorbent assay（ELISA） method was used to detect the titer of the polyclonal antibodies； Western blotting method was used to verify the specificity and sensitivity of the antibodies and to determine the natural expression of the protein； immunofluorescence assay （IFA） was used to analyze the subcellular localization of the protein. Results The bioinformatics analysis results showed that Trx20 protein was a relatively stable hydrophilic protein with a molecular formula of C₂₁₇₂H₃₄₁₂N₅₄₈O₆₁₆S₂₀， containing 424 amino acids， a predicted relative molecular mass of 47 700， and a theoretical isoelectric point of 8.55； it was predicted that the protein had one signal peptide， no transmembrane region， contained one domain named “Thioredoxin like Superfamily”， and had 35 phosphorylation sites， one N-glycosylation site， and 17 antigenic determinants； in the secondary structure， alpha-helices accounted for 41.51% of the total amino acids， and random coils accounted for 39.86%； the recombinant plasmids pET-28a-Trx20 and pGEX-4T-1-Trx20 were successfully constructed， and the soluble recombinant protein was expressed and purified； polyclonal antibodies were successfully prepared with a titer as high as 1∶64 000， and they specifically recognized the endogenous Trx20 protein in Toxoplasma gondii； the subcellular localization results showed that Trx20 protein was widely distributed in the cytoplasm of the parasite. Conclusion Toxoplasma gondii Trx20 protein is a secretory protein containing phosphorylation/glycosylation modification sites and a thioredoxin domain， and it is localized in the cytoplasm of the parasite.

Objective To discuss the effect of berberine hydrochloride （BBR） on autophagy in the HeLa cells infected with herpes simplex virus type 1 （HSV-1）， and to clarify its related mechanism. Methods The HeLa cells at logarithmic growth phase were added with 0， 10， 20， 40， 80， 120， and 160 μmol·L^-1 BBR， respectively， and cultured for 24 h. In addition， the HeLa cells were divided into HSV-1 group （the cells were infected with HSV-1）， L-BBR group （were infected with HSV-1 and then treated with 20 μmol·L^-1 BBR for 24 h）， M-BBR group （were infected with HSV-1 and then treated with 40 μmol·L^-1 BBR for 24 h）， H-BBR group （were infected with HSV-1 and then treated with 80 μmol·L^-1 BBR for 24 h）， and 740 Y-P group （were infected with HSV-1 and then treated with 80 μmol·L^-1 BBR and 10 μmol·L^-1 740 Y-P for 24 h） for viral infection and corresponding drug treatment. MTT method was used to detect the activities of microtubule-associated protein 1 light chain 3 （LC3） the HeLa cells after treated with different concentrations of BBR； plaque reduction assay was used to detect the proliferation ability of HSV-1 in the HeLa cells in various groups； real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the mRNA expression levels of virus replication-related genes in the HeLa cells in various groups； immunofluorescence method was used to detect the formation of autophagosomes in the HeLa cells in various groups； flow cytometry was used to detect the apoptotic rate of the HeLa cells in various groups； Western blotting method was used to detect the expression levels of autophagy and the phosphoinositide 3-kinase （PI3K）/protein kinase B（AKT）/mammalian target of rapamycin （mTOR） pathway pathway-related proteins in the HeLa cells in various groups. Results The MTT method results showed that compared with 0 μmol·L^-1 BBR group， the activities of the HeLa cells in 120 and 160 μmol·L^-1 BBR groups were significantly decreased （P<0.05）， which had certain toxicity to the cells； 20， 40， and 80 μmol·L^-1 BBR were selected for subsequent experiments. The plaque reduction assay results showed that compared with HSV-1 group， the plaque forming units（PFUs） in the HeLa cells in L-BBR group， M-BBR group， and H-BBR group were significantly decreased （P<0.05） in a dose-dependent manner； compared with H-BBR group， the PFUs in the HeLa cells in 740 Y-P group were significantly increased （P<0.05）. The RT-qPCR results showed that compared with HSV-1 group， the mRNA expression levels of infected cell protein 0 （ICP0）， infected cell protein 22 （ICP22）， infected cell protein 8 （ICP8）， thymidine kinase （TK）， glycoprotein B （gB）， and glycoprotein D （gD） in the HeLa cells in L-BBR group， M-BBR group， and H-BBR group were significantly decreased （P<0.05） in a dose-dependent manner； compared with H-BBR group， the expression levels of ICP0， ICP22， ICP8， TK， gB， and gD mRNA in the HeLa cells in 740 Y-P group were significantly increased （P<0.05）. The immunofluorescence method results showed that compared with HSV-1 group， the number of LC3 autophagosomes formed in the HeLa cells in L-BBR group， M-BBR group， and H-BBR group was increased； compared with H-BBR group， the number of LC3 autophagosomes formed in the HeLa cells in 740 Y-P group was decreased. The flow cytometry results showed that compared with HSV-1 group， the apoptotic rates of the HeLa cells in L-BBR group， M-BBR group， and H-BBR group were significantly increased （P<0.05） in a dose-dependent manner； compared with H-BBR group， the apoptotic rate of the HeLa cells in 740 Y-P group was significantly decreased （P<0.05）. The Western blotting method results showed that compared with HSV-1 group， the expression levels of Beclin-1 and B-cell lymphoma 2 （Bcl-2）/adenovirus E1B 19kDa protein interacting protein protein（BNIP） 2 proteins and the ratios of LC3-Ⅱ/LC3-Ⅰ in the HeLa cells in L-BBR group， M-BBR group， and H-BBR group were significantly increased （P<0.05） in a dose-dependent manner； compared with H-BBR group， the expression levels of Beclin-1 and BNIP proteins and the LC3-Ⅱ/LC3-Ⅰ ratio in the HeLa cells in 740 Y-P group were significantly decreased （P<0.05）； compared with HSV-1 group， the expression levels of cysteine-containing aspartate protein hydrolase 1 （Caspase-1）， cysteine-containing aspartate protein hydrolase 3 （Caspase-3）， and Bcl-2 associated X protein （Bax） proteins in the HeLa cells in L-BBR group， M-BBR group， and H-BBR group were significantly increased （P<0.05）， and the expression level of Bcl-2 protein was significantly decreased （P<0.05） in a dose-dependent manner； compared with H-BBR group， the expression levels of Caspase-1， Caspase-3， and Bax proteins in the HeLa cells in 740 Y-P group were significantly decreased （P<0.05）， and the expression level of Bcl-2 protein was significantly increased （P<0.05）； compared with HSV-1 group， the ratios of phosphorylated PI3K （p-PI3K）/PI3K， phosphorylated AKT（p-AKT）/AKT， and phosphorylated mTOR （p-mTOR）/mTOR in the HeLa cells in L-BBR group， M-BBR group， and H-BBR group were significantly decreased （P<0.05） in a dose-dependent manner； compared with H-BBR group， the ratios of p-PI3K/PI3K， p-Akt/Akt， and p-mTOR/mTOR in the HeLa cells in 740 Y-P group were significantly increased （P<0.05）. Conclusion BBR can promote the autophagy process in the HeLa cells infected with HSV-1 by regulating the PI3K/AKT/mTOR pathway， induce autophagy-dependent apoptosis， and significantly inhibit the virus replication.

Objective To discuss the expression and clinical significance of inositol polyphosphate-4-phosphatase type Ⅱ （INPP4B） gene in hepatocellular carcinoma （HCC） based on The Cancer Genome Atlas （TCGA） database and experimental verification with clinical samples. Methods Based on data from 424 clinical samples in the TCGA database （including 374 HCC tissues and 50 paracarcinoma tissues）， Kaplan-Meier method and Cox regression analysis were used to evaluate the relationship between INPP4B gene and the clinical characteristics and survival prognosis of the HCC patients. The correlations between INPP4B gene and the number of 24 types of immune cells， matrix， immune cell infiltration and tumor purity in tumor tissue， and the expression level of the high-frequency mutant gene tumor protein 53（TP53） in HCC were analyzed. The clinicopathological data and paraffin-embedded tissue sections of 60 HCC patients treated with surgical resection from December 2022 to December 2023 were collected. According to clinical diagnosis， they were divided into poorly differentiated group （HCC-L group）， moderately differentiated group （HCC-M group） and well-differentiated group （HCC-H group）， with 20 cases in each group； 20 patients during the same period who underwent biopsy and were pathologically diagnosed as non-tumor were selected as normal group， and their clinicopathologic data and liver tissue paraffin sections were collected. HE staining was used to observe the pathomorphology of HCC tissue and normal liver tissue of the subjects in various groups； immunohistochemistry method was used to detect the expressions of Ki-67 and INPP4B proteins in the HCC tissue and normal liver tissue of the subjects in various groups. Results The TCGA database analysis results showed that compared with normal tissue， the expression level of INPP4B mRNA in HCC tissue was significantly increased （P<0.01）. Compared with INPP4B low expression group， the overall survival（OS） of the patients in INPP4B high expression group was significantly prolonged （P<0.05）. The univariate Cox regression analysis results showed that tumor stage， pathological stage， tumor status and residual tumor had impacts on OS of the HCC patients （P<0.05）. The univariate regression analysis results showed that the INPP4B prognostic risk model score ratio was HR=0.781， 95% confidence interval（CI）： 0.552-1.105， P=0.168. The AUC value for the impact of INPP4B on OS of the HCC patients was 0.558， indicating that the INPP4B gene prognostic risk model had certain predictive value in survival prognosis. The INPP4B mRNA expression level was not correlated with TNM stage， stage， patient gender， age， race or body mass index （BMI） （P>0.05）. In tumor tissue with high and low INPP4B expression， 22 types of immune cells showed statistically significant differences （P<0.05）； the INPP4B mRNA expression level was positively correlated with the number of 23 types of immune cells except T helper （Th） 17 cells （r>0）， among which all Th cells except natural killer （NK） CD56⁺ cells were statistically significant （P<0.01）； INPP4B was significantly correlated with matrix （r=0.475）， immune cell infiltration （r=0.641） and tumor purity （r=0.599） in tumor tissue （P<0.01）. INPP4B was correlated with TP53 （r=0.287， P<0.01）. The HE staining results showed that clear and complete lobular structure， neatly arranged cells and slight inflammatory cell infiltration were observed in liver tissue of the subjects in normal group； completely destroyed lobular structure， significant hepatocellular steatosis， massive inflammatory cell infiltration， and lesions such as ballooning degeneration and small cell hyperplasia in some cells were observed in HCC tissue of the patients in HCC-L， HCC-M and HCC-H groups， and the lower the HCC differentiation degree， the more severe the tissue destruction； The immunohistochemistry results showed that compared with normal group， the expression levels of Ki-67 protein in HCC tissue of the patients in HCC-L， HCC-M and HCC-H groups were significantly increased （P<0.01）， and the lower the differentiation degree of the HCC patients， the higher the Ki-67 positive rate. Brownish-yellow granules evenly distributed in the cells and INPP4B protein was highly expressed in liver tissue of the subjects in normal group； compared with normal group， the expression levels of INPP4B protein in HCC tissue of the patients in HCC-L， HCC-M and HCC-H groups were significantly decreased （P<0.01）， and the lower the differentiation degree of the HCC tissue， the lower the INPP4B positive rate. Conclusion INPP4B is a protective factor for the prognosis of HCC patients； as a new tumor suppressor gene， INPP4B may become a potential target for new drug screening in HCC treatment.

Objective To discuss the clinical characteristics of the patients with hypertrophic cardiomyopathy （HCM） complicated with microcirculatory dysfunction （CMD）， and to analyze the impact of concurrent CMD on the prognosis of the HCM patients. Methods A total of 211 patients diagnosed with HCM and having complete cardiac magnetic resonance imaging （CMR） examination results from January 1， 2019 to September 30， 2023 were collected. They were divided into HCM complicated with CMD group （68 cases） and HCM complicated without CMD group （143 cases） based on CMR assessment. The clinical data such as age， gender， admission symptoms， and past medical history， blood test data such as troponin， electrocardiogram， echocardiography， and CMR data including abnormal Q wave， ST segment depression， inverted T wave， PR interval， QRS wavelength， corrected QT interval， ejection fraction （EF）， left atrial diameter （LAD）， left and right ventricular end-diastolic diameters， cardiac output， E peak， A peak， and maximum wall thickness （MWT） of the patients were compared between two groups. Logistic regression was used to analyze the clinical characteristics of the HCM patients complicated with CMD； multivariate modified Poisson regression was used to analyze the risk factors for major adverse cardiovascular events （MACE） in the HCM patients. Results Compared with HCM complicated without CMD group， the percentage of palpitation patients in HCM complicated with CMD group was significantly increased （P<0.05）， the percentage of tachycardia episode patients was significantly increased （P<0.05）， the troponin level was significantly increased （P<0.05）， and the percentage with a history of hypertension patients was significantly decreased （P<0.05）. Compared with HCM complicated without CMD group， the percentage of abnormal Q wave on electrocardiogram in the patients in HCM complicated with CMD group were significantly increased （P<0.05）， the percentage of inverted T wave and the EF of the patients was significantly decreased （P<0.05）， the LAD was significantly increased （P<0.05）， and the MWT was significantly increased （P<0.05）. The multivariate Logistic regression analysis results showed that increased LAD （OR=1.05， 95%CI： 1.00-1.11， P=0.048） and increased MWT （OR=1.11， 95%CI： 1.03-1.19， P=0.007） were the risk factors for concurrent CMD in the HCM patients； history of hypertension （OR=0.40， 95%CI： 0.20-0.80， P=0.010） was a protective factor for concurrent CMD in the HCM patients. The average follow-up time in this study was 20.5 months. A total of 27 patients experienced MACE， with an overall incidence of 12.80%， including 12 patients in HCM complicated with CMD group and 15 patients in HCM complicated without CMD group. The multivariate modified Poisson regression analysis results showed that history of diabetes （RR=2.34， 95%CI： 1.09-5.06， P=0.030）， history of arrhythmia （RR）=4.00， 95%CI： 1.82-8.83， P=0.001）， and decreased ejection fraction （RR=0.96， 95%CI： 0.94-0.99， P=0.001） were risk factors for MACE in the HCM patients. Conclusion The HCM patients complicated with CMD have unique clinical characteristics， including higher symptom burden， left atrial enlargement， myocardial hypertrophy， and increased troponin levels. Concurrent CMD does not increase the short-term risk of adverse events； diabetes， arrhythmia， and decreased EF are key risk factors for prognosis； early intervention and complication management for HCM complicated with CMD patients may improve the long-term prognosis of the HCM patients.

Objective To explore the expressions of calcium-activated chloride channel protein 1（ANO1） and E-cadherin in colorectal cancer （CRC） tissue and their relationships with the clinicopathological characteristics of the CRC patients， and to clarify their clinical significances. Methods The surgical tissue specimens from 77 patients with primary CRC were collected. Immunohistochemistry （IHC） method was used to detect the protein expressions of ANO1 and E-cadherin in CRC tissue； Spearman correlation analysis method was used to analyze the correlation between their expressions； PCR fluorescence probe method was used to detect the KRAS/NRAS mutations in CRC tissue； chi-square test or Fisher’s exact probability method was used to analyze the relationships between the expressions of ANO1 and E-cadherin and clinicopathological characteristics of the CRC patients. Results The IHC results showed that the positive expression rate of ANO1 in CRC tissue was 24.7%， and the positive expression rate of E-cadherin was 63.6%. The Spearman analysis results showed that the expressions of ANO1 and E-cadherin in CRC tissue were positively correlated （r=0.458， P<0.05）. Compared with the patients with tumor located in rectum， the positive expression rate of ANO1 in cancer tissue of the CRC patients with tumor located in colon was significantly increased （χ²=5.499， P=0.019）； compared with the patients with TNM stage Ⅲ-Ⅳ， the positive expression rate of ANO1 in cancer tissue of the CRC patients with TNM stage Ⅰ-Ⅱ was significantly increased （χ²=4.774， P=0.029）； compared with the patients with lymphnode metastasis， the positive expression rate of ANO1 in the CRC patients without lymphnode metastasis was significantly increased （P=0.034）. Compared with the CRC patients with T4 stage， the positive expression rate of E-cadherin in the CRC patients with T1-T3 stage was significantly increased （P=0.024）. Compared with the CRC patients with p53 positive， Ki-67≥90%， PMS2 positive and KRAS wild-type， the positive expression rate of ANO1 in cancer tissue of the CRC patients with p53 negative， Ki-67<90%， PMS2 negative and KRAS mutant was significantly increased （P=0.031， P=0.036， P=0.048， P=0.028）. Conclusion The expressions of ANO1 and E-cadherin in primary CRC tissue of the patients are positively correlated； ANO1 and E-cadherin are associated with multiple clinicopathological characteristics of the patients； ANO1 and E-cadherin may be involved in the occurrence and progression of CRC.

Objective To discuss the clinical efficacy of microwave ablation （MWA） and surgical treatment in the patients with Hashimoto thyroiditis （HT） complicated with T1aN0M0 stage papillary thyroid carcinoma （PTC）， and to clarify the feasibility and advantages of MWA as a minimally invasive treatment modality. Methods A retrospective cohort study was used. A total of 165 patients diagnosed with HT complicated with T1aN0M0 stage PTC from January 2018 to December 2020 were selected and divided into MWA group （82 cases） and surgery group （83 cases）. The patients in MWA group underwent MWA treatment under ultrasound guidance， and the patients in surgery group underwent unilateral thyroid lobectomy + unilateral lymph node dissection. The median follow-up time was （65.98±7.32） months. The indicators such as age， preoperative tumor volume， operation time， intraoperative blood loss， hospital stay， hospitalization cost， tumor progression， incidence of postoperative complications， changes in thyroid function， tumor recurrence rate， and life quality score of the patients in two groups were recorded and analyzed. Results There were no significant differences in the baseline sample data including age， gender， preoperative tumor volume， margin， shape， aspect ratio， calcification， and preoperative thyroid function between two groups （P>0.05）. Compared with surgery group， the operation time， intraoperative blood loss， hospital stay， and hospitalization cost of the patients in MWA group were significantly decreased （P<0.01）， and there was no significant difference in the incidence of postoperative complications （P>0.05）. All nodules in the patients in MWA group achieved complete ablation within 18 months. The functional thyroid parenchyma in the patients in MWA group was well preserved after operation， and the thyroid function was maintained within the normal physiological range during the follow-up period. The Thyroid Cancer-specific Quality of Life Questionnaire（THYCA-QoL） results showed that compared with MWA group， the scores of voice， sympathetic nerve symptoms， throat/mouth problems， psychological， sensory problems， scar， and chills of the patients in surgery group were significantly inecreased （P<0.05）. There was no significant difference in the tumor progression rate between two groups （P>0.05）. Conclusion During the median follow-up period， both MWA and surgical treatment achieve satisfactory clinical efficacy in the patients with HT complicated with T1aN0M0 stage PTC. MWA treatment has significant advantages in operation time， intraoperative blood loss， hospital stay， hospitalization cost， quality of life， and preservation of thyroid function， and it can be used as a minimally invasive treatment option for such patients， but its long-term oncological safety needs to be further evaluated by extending the follow-up.

Objective To discuss the clinical value of pan-immune inflammation index （PIV） in predicting the major adverse cardiovascular events （MACE） within 1 year after percutaneous coronary intervention （PCI） in the elderly patients with coronary heart disease， and to clarify the role of inflammatory response in postoperative recovery and prognosis of the patients with coronary heart disease. Methods A total of 150 elderly patients with coronary heart disease who underwent PCI from July 2020 to August 2023 were selected as the research subjects； according to the occurrence of MACE within 1 year after operation， they were divided into MACE group （n=28） and non-MACE group （n=122）； the baseline data and biochemical indicators of the patients were collected， and PIV was calculated； multivariate Logistic regression was used to analyze the influencing factors of MACE within 1 year after PCI in the elderly patients with coronary heart disease； receiver operating characteristic （ROC） curve was used to analyze the predictive value of PIV for MACE within 1 year after PCI in the elderly patients with coronary heart disease. Results Compared with non-MACE group， the levels of total cholesterol （TC） and low-density lipoprotein cholesterol （LDL-C）， neutrophils （NEUT）， platelets （PLT） counting and PIV in the patients in MACE group were significantly increased （P<0.05）； there were no significant differences in other data between two groups （P>0.05）. The multivariate Logistic regression analysis results showed that the levels of TC （OR=1.571， 95%CI： 1.088-2.270） and LDL-C （OR=32.506， 95%CI： 8.880-118.994） and PIV （OR=1.014， 95%CI： 1.010-1.019） were the influencing factors of MACE within 1 year after PCI in the elderly patients with coronary heart disease （P<0.05）. The ROC curve analysis results showed that the area under the ROC curve （AUC） of PIV for predicting MACE was 0.857 （95%CI： 0.762-0.951）， the sensitivity was 0.821， the specificity was 0.959， the maximum Youden index was 0.780， and the best cut-off value was 778.805 （P<0.01）. Conclusion PIV has important predictive value for MACE within 1 year after PCI in elderly patients with coronary heart disease.

Objective To discuss the differential expression of microRNA（miR）-125a-5p in peripheral blood mononuclear cells （PBMCs） of the patients with mycobacterium tuberculosis （MTB） infection and its effect on macrophage polarization， and to clarify its clinical significance. Methods A total of 40 patients with active tuberculosis （ATB） （ATB group）， 35 patients with latent tuberculosis infection （LTBI） （LTBI group）， and 40 healthy physical examinees （control group） clinically diagnosed from July 2022 to June 2023 were selected. The fasting blood samples of the subjects in three groups were collected next morning after 12 h of fasting， and then serum was separated. Enzyme-linked immunosorbent assay（ELISA） method was used to detect the levels of inflammatory factors in the serum of the subjects in various groups. Simultaneously， the PBMCs were extracted from the subjects in various groups； flow cytometry was used to detect the expression levels of CD80 and CD206 proteins in the PBMCs of the subjects in various groups； real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of microRNA （miR）-125a-5p and interleukin-6 （IL-6） mRNA in PBMCs of the subjects in various groups. Results There were no statistically significant differences in the general information of the subjects among three groups （P>0.05）. Compared with control group， the erythrocyte sedimentation rate （ESR）， percentages of monocytes （MONO）， tumor necrosis factor-α （TNF-α） levels， and interleukin-10 （IL-10） levels in serum of the patients in ATB group and LTBI group were significantly increased （P<0.05）， and the percentages of lymphocytes （LY） were significantly decreased （P<0.05）； compared with ATB group， the ESR and level of IL-10 in serum of the patients in LTBI group were significantly decreased （P<0.05）， and the percentage of LY was significantly increased （P<0.05）； there were statistically significant differences in the counts of white blood cell （WBC） of the subjects among various groups （P>0.05）. The flow cytometry results showed that compared with control group， the expression levels of CD80 and CD206 proteins in PBMCs of the patients in ATB group and LTBI group were significantly increased （P<0.05）. Compared with ATB group， the expression level of CD206 protein in the PBMCs of the patients in LTBI group was significantly increased （P<0.05）， and the expression level of CD80 protein was significantly decreased （P<0.05）. The RT-qPCR results showed that compared with control group， the expression levels of miR-125a-5p and IL-6 mRNA in the PBMCs of the patients in ATB group and LTBI group were significantly increased （P<0.05）； compared with ATB group， the expression levels of miR-125a-5p and IL-6 mRNA in PBMCs of the patients in LTBI group were significantly increased （P<0.05）. The correlation analysis results showed that the miR-125a-5p expression level was positively correlated with the TNF-α level and IL-6 mRNA expression level （r=0.406， P<0.05； r=0.351， P<0.05）， and negatively correlated with the IL-10 level （r=-0.368， P<0.05）. The area under the receiver operating characteristic （ROC） curve （AUC） value of miR-125a-5p expression level for diagnosing LTBI patients was 0.89 （P<0.01）， with a sensitivity of 0.85 and a specificity of 0.88. Conclusion The expression level of miR-125a-5p in PBMCs of the patients in LTBI group is significantly increased， and it can affect the macrophage polarization to M1， promote the inflammatory response process of macrophages and participate in the occurrence and development of pulmonary tuberculosis.

Objective To analyze the breast cancer-related parameters and the ultrasonic features of positive axillary lymph nodes， to discuss the risk factors for axillary lymphnode metastatic burden， and to provide basis for preoperative evaluation of breast cancer patients. Methods The ultrasonic and clinicopathological data of 574 breast cancer patients with axillary lymph node metastasis confirmed by surgery and pathology were retrospectively analyzed. According to the status of axillary lymphnode metastasis， the patients were divided into low nodal burden （LNB） group （n=283） and high nodal burden （HNB） group （n=291）. The affected side， tumor quadrant， distance to skin， maximum diameter， internal echogenicity， shape， margin， calcification， blood supply， posterior echo， lymphnode long diameter， lymphnode short diameter， lymphnode aspect ratio， number of suspicious metastases， intranodal blood supply， lymphnode hilum morphology， age， pathological type， histological grade， molecular subtype， and the expressions of estrogen receptor （ER）， progesterone receptor （PR）， Ki-67， human epidermal growth factor receptor 2 （HER2）， and P53 were compared between two groups. Logistic regression was used to analyze the risk factors for axillary lymph node metastatic burden in the breast cancer patients； receiver operating characteristic （ROC） curve and area under the curve （AUC） were used to evaluate the predictive value. Results The univariate analysis results showed that there were statistically significant differences in tumor quadrant， distance to skin， molecular subtype， HER2 positive expression， lymphnode long diameter， lymph node short diameter， lymph node aspect ratio， number of suspicious metastases， and lymphnode hilum morphology between two groups （P<0.05）. The multivariate Logistic regression analysis results showed that tumor located in the upper outer quadrant （OR=0.648， P=0.021）， distance to skin <5 mm （OR=0.283， P=0.016）， Luminal A （OR=1.564， P=0.044）， lymphnode long diameter ≥20 mm （OR=2.050， P<0.01）， lymphnode short diameter ≥8.6 mm （OR=2.430， P<0.01）， lymph node aspect ratio <2 （OR=1.585， P<0.01）， and indistinct lymphnode hilum structure （OR=2.092， P<0.01） were the independent risk factors for axillary lymphnode metastatic burden. The ROC curve analysis results showed that compared with the ultrasonic features of positive axillary lymph nodes， the AUC of the combination of breast cancer-related parameters and ultrasonic features of positive axillary lymphnodes was larger （Z=2.72， P=0.006 5）， and it had higher predictive value for axillary lymphnode metastatic burden. Conclusion The tumor quadrant， distance to skin， molecular subtype， lymphnode long diameter， lymph node short diameter， lymphnode aspect ratio， and lymphnode hilum structure are the independent risk factors for axillary lymphnode metastatic burden， and they have certain predictive value for axillary lymphnode metastatic burden.

Objective To explore the clinical application effect of belimumab in the treatment of systemic lupus erythematosus （SLE） and its influence on cluster of differentiation （CD40） and cluster of differentiation 40 ligands （CD40L） expression in the peripheral blood， and to provide a theoretical basis for the effective treatment of SLE. Methods A retrospective analysis was conducted on the clinical data of 150 patients with SLE admitted to our hospital， and they were divided into conventional treatment group （n=78， standard treatment plan） and belimumab group （n=72， standard treatment plan + belimumab） according to the treatment methods. The therapeutic effects and adverse reactions were compared. The clinical symptoms， organ involvement， disease activities， prednisone doses， the expression levels of CD40 and CD40L mRNA in peripheral blood mononuclear cells， and the levels of immunology-related indicators before and after treatment were analyzed. Results Compared with conventional treatment group， the total effective rate， complement C3 and complement C4 levels in the belimumab group were significantly increased after treatment （P<0.01 or P<0.001）， while the proportion of renal involvement， the SLE disease activity index-2K （SLEDAI-2K） score， the dosage of prednisone， immunoglobulin G （IgG） level and the expression levels of CD40 and CD40L mRNA in peripheral blood mononuclear cells were decreased （P<0.01 or P<0.001）. There was no statistically significant difference in the total incidence of adverse reactions between two groups during treatment period （P>0.05）. Conclusion Belimumab can increase the effective rate of SLE treatment， reduce disease activity， decrease the involvement of the kidneys and the dose of hormones， and has good safety； its mechanism of action may be related to down-regulating the expression of CD40 and CD40L in peripheral blood and improving the immune function.

Limited-stage small cell lung cancer （SCLC） is a highly malignant and rapidly progressing neuroendocrine tumor， while uremia is a complication of the end-stage of chronic renal failure. The patients with SCLC complicated with uremia have poor treatment tolerance， limited options for anti-tumor treatment regimens， and great difficulty in diagnosis and treatment. This study analyzed one case of a 69-year-old male patient with limited-stage SCLC complicated with uremia （with a history of regular hemodialysis， 3 times per week）， to discuss his first-line treatment regimen， efficacy， and the impact of hemodialysis on the plasma concentrations of the anti-tumor drugs， and reviewed the relevant literature to provide a reference for the treatment of similar patients. The patient was admitted to the hospital due to “cough and hemoptysis for half a month” and was diagnosed with limited-stage SCLC stage ⅢA （T2aN2M0） by computed tomography （CT） and lung puncture biopsy. After discussion by the multi-disciplinary treatment （MDT） team， the patient received 6 cycles of Etoposide（VP-16） + carboplatin chemotherapy combined with adebrelimab immunotherapy， followed by sequential adebrelimab maintenance therapy. The efficacy was evaluated as partial response （PR） and the response is ongoing. During the treatment， level 4 hemoglobin decrease， level 3 neutropenia， and level 2 leukopenia occurred， which were alleviated after symptomatic treatment. The blood concentration monitoring results showed that the plasma concentrations of etoposide and carboplatin increased rapidly during drug infusion， and gradually decreased after the end of infusion. Hemodialysis could rapidly reduce the plasma concentration of carboplatin， but had no significant effect on the plasma concentration of etoposide. Therefore， the immunotherapy combined with reduced-dose chemotherapy regimen is safe and effective for this type of patient. Plasma drug concentration monitoring can be used to observe drug metabolism， but the optimal monitoring time points and clinical value need further study and validation.

Follicular variant of papillary thyroid carcinoma （FVPTC） is commonly found in young and middle-aged women， and cases in children and adolescents are relatively rare. The main driving genes of FVPTC are the RAS oncogene family or B-Raf proto-oncogene serine/threonine protein kinase （BRAF）， and DICER1 gene mutation is rarely reported in this subtype. This article reports a case of recurrent FVPTC with positive DICER1 mutation in an adolescent， aiming to provide reference for the clinical diagnosis and treatment of FVPTC. The patient， a 19-year-old female， underwent total thyroidectomy and central lymph node dissection due to thyroid nodules 5 years ago， and was pathologically diagnosed with follicular tumor of uncertain malignant potential （FT-UMP）； after surgery， the patients did not regularly undergo thyroid-stimulating hormone （TSH） suppression therapy and follow-up as instructed. Two months ago， the patient visited the hospital due to enlarged cervical lymph nodes， imaging suggested tumor recurrence with metastases to both lungs and vertebrae， and multigene testing indicated a DICER1 gene mutation. After comprehensive multidisciplinary consultation， bilateral cervical lymph node dissection was performed， and postoperative pathology confirmed the diagnosis of metastatic FVPTC. Te clinicans should focus on rare gene mutations such as DICER1 in the FVPTC patients， and for adolescent patients， follow-up and formulation of individualized treatment plans should be emphasized to achieve early tumor identification and precise intervention， and improve the long-term prognosis of patients.

Objective To construct the sphingosine kinase 1 （SPHK1） overexpression lentiviral vector， and to establish the SKOV3 lentiviral stable transfection cell line. Methods According to the SPHK1 data information provided by the National Center for Biotechnology Information （NCBI） database， the primers were designed and synthesized， the target gene was amplified， and connected to the GV492 plasmid treated with BamHⅠ and AgeⅠ restriction enzymes to construct the SPHK1 overexpression lentiviral vector； the positive clones were selected for PCR and sequencing identification； the lentiviral plasmid and the lentiviral packaging auxiliary plasmid were co-transfected into the HEK-293T cells for packaging and titer determination； according to the measured optimal multiplicity of infection （MOI） of 10， the corresponding lentiviral amounts in various groups were transfected into the SKOV3 cells， and the SKOV3 cells were divided into blank group （without treatment）， GV492 control group （GV492 control lentivirus infected SKOV3 cells）， and GV492-SPHK1 overexpression group （GV492-SPHK1 overexpression lentivirus infected SKOV3 cells， ov-SPHK1 group）； the optimal concentration of 2 mg·L^-1 puromycin was used to screen the stably transfected SKOV3 cell line； after 48 h， the medium was changed and replaced with 1 mg·L^-1 puromycin for screening for 14 d； the morphology and fluorescence expression of the cells were observed under fluorescence microscope； real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of SPHK1 mRNA in the SKOV3 cells in various groups； Western blotting method was used to detect the expression level of SPHK1 protein in the SKOV3 cells in various groups. Results The PCR sequencing results showed that the gene sequence of the SPHK1 overexpression lentiviral vector was completely consistent with the target sequence， and the SPHK1 overexpression lentiviral vector was successfully constructed； the titer determination results showed that the lentiviral titers in GV492 control group and ov-SPHK1 group were 5×1011 and 8×1011 TU·L?1， respectively； the SKOV3 cells in GV492 control group and ov-SPHK1 group were in good state and showed strong fluorescence expression， suggesting that the SKOV3 stable transfection cell line overexpressing SPHK1 was successfully established； the RT-qPCR results showed that compared with blank group and GV492 control group， the expression level of SPHK1 mRNA in the SKOV3 cells in ov-SPHK1 group was significantly increased （P<0.01）； the Western blotting results showed that compared with blank group and GV492 control group， the expression level of SPHK1 protein in the SKOV3 cells in ov-SPHK1 group was significantly increased （P<0.01）. Conclusion The SPHK1 overexpression lentiviral vector is successfully constructed， and the SKOV3 stable transfection cell line is established.

Objective To construct the prokaryotic expression vector of Coxsackievirus A16 （CA16） viral protein 1 （Vp1）， express it in Escherichia coli （E.coli） BL21， purify the protein， prepare rabbit polyclonal antibodies， and identify the immunoreactivity of the antibodies. Methods Bioinformatics online tools were used to predict the amino acid composition， conserved domains， secondary and tertiary structures of the Vp1 protein. The Vp1 gene of CA16 was amplified and cloned into the prokaryotic expression vector pET28a（+）. The pET28a-Vp1 was transformed into E.coli BL21， and the expression was induced using isopropyl β-D-thiogalactopyranoside （IPTG）. Western blotting method was used to identify the induced expression of Vp1 protein， and the induction time and temperature were optimized to improve the expression efficiency. Two female SPF New Zealand white rabbits were taken， and the purified recombinant protein Vp1 was used to immunize the rabbits to prepare rabbit anti-Vp1 protein polyclonal antibodies. The antibody titer was determined by ELISA method， and the immunoreactivity of the antibodies was identified by Western blotting method. Results The bioinformatics analysis results showed that the Vp1 gene encoded 297 amino acids， with a relative molecular mass of 33 046.39 and an isoelectric point of 8.32， belonging to a hydrophilic protein； in the secondary structure， α-helix accounted for 15.15%， random coil accounted for 67.68%， and extended strand accounted for 17.17%. Sequencing of the recombinant plasmid pET28a-Vp1 confirmed that the pET28a-Vp1 plasmid was correctly constructed. The Western blotting results showed that the target protein was expressed in the IPTG induction group at a relative molecular mass of 33 000， and the target protein was mainly expressed in inclusion bodies. The optimal induction conditions for protein expression were IPTG concentration of 0.4 mmol·L^-1， temperature of 16 ℃， and induction time of 20 h. The ELISA assay results showed that the titer of the rabbit polyclonal antibody was 1∶1 024 000. The Western blotting results showed that the Vp1 rabbit polyclonal antibody could bind to the viral Vp1 protein in the CA16-infected cells. Conclusion The polyclonal antibody against CA16 Vp1 is successfully prepared， and this antibody has significant binding characteristics to CA16 Vp1， which can be used for the diagnosis of enterovirus infection and the development of treatment methods.

Emotional dysregulation （ED） is a common problem in the children with autism spectrum disorder （ASD） and has adverse effects on social interaction， academic achievement， parent-child relationships， and overall functioning. The comorbid ED in the children with ASD is easily confused with or overlooked due to its core symptoms. Assessing the emotional regulation （ER） problems in the children with ASD from multiple perspectives is helpful for the early identification and diagnosis of ED， and can provide the reference for developing intervention methods to improve ED and even the overall prognosis of the children with ASD. Discussing the application of non-pharmacological and pharmacological treatment methods for ED in the children with ASD is helpful for selecting appropriate intervention strategies for ED in different subgroups of the children with ASD， and can also provide the basis for intervention methods more suitable for ED in toddler and preschool-aged children with ASD. Based on relevant research results at home and abroad， this article introduces the research progress in comorbid ED in the children with ASD from the aspects of the current status of emotional dysregulation， assessment methods， and intervention strategies， in order to provide more scientific basis for the early identification， diagnosis， and effective intervention of ED in children with ASD.

RNA-cleaving deoxyribozymes （DNAzymes） are the DNA molecules with catalytic RNA cleavage activity， abd they can cleave the RNA substrates with high specificity with the advantages such as high specificity， no permanent impact on the genome， simple design and sequence- programmable， giving them great potential in biosensing and disease therapy. However， off-target activation of deoxyribozyme in vivo or complex biological environments limits their sensitivity and selectivity in various applications. Recent studies have focused on the chemical modification of deoxyribozymes， precise control for their catalytic activity its performed by introducing specific chemical groups； in addition， by combining deoxyribonucleases with aptamers， their precise recognition and efficient cleavage of target molecules can be realized； and the deoxyribozyme sequences can be used for logical operations and the development of molecular computers after suitable design. In this review， various approaches to the modification and design of DNAzymes were comprehensively discussed， with their unique advantages and superiority in biosensing and gene therapy， including examples of chemically and structurally modified RNA-cleaving depxurobozyme for therapeutic and sensing applications in the recent years.Ultimately， the review discussed the challenges and prospects of RNA-cleaving DNAzymeby for its applications in disease diagnostic and therapy， aiming to provide the reference farthe further application of deoxyribozyme in related fields.

Endometrial cancer is one of the common malignant tumors of the female reproductive system. Its incidence rate in China is second only to cervical cancer， posing a serious threat to women’s life and health， and the onset shows a trend of affecting younger individuals. Currently， the primary treatment for endometrial cancer is surgical intervention， and some postoperative patients and those with postoperative recurrence require adjuvant radiotherapy. Pelvic external beam radiotherapy（EBRT） and vaginal brachytherapy（VBT） are two modalities of postoperative radiotherapy. The appropriate postoperative radiotherapy approach should be selected based on individual conditions and guideline recommendations. Most patients can achieve satisfactory treatment outcomes through surgery and postoperative adjuvant radiotherapy. However， the specific indications for postoperative radiotherapy in endometrial cancer patients and the radiotherapy principles for those with postoperative recurrence are relatively complex， and recommendations for radiotherapy modalities in postoperative patients vary slightly across different guidelines. This article reviews the treatment principles for endometrial cancer patients in recent years， as well as the indications for postoperative and post operative recurrent radiotherapy and the radiotherapy techniques， aiming to provide a reference for radiotherapy in the endometrial cancer patients.

Skin diseases significantly affect the quality of life of approximately 190 million individuals worldwide. The complexity and diversity of their clinical manifestations are the major challenges for traditional diagnostic approaches， and exploring novel diagnostic strategies has become an urgent priority. In recent years， deep learning （DL） technology has been increasingly applied in the intelligent recognition of skin diseases， demonstrating substantial potential. This study provides a systematic review of the research progress of DL in dermatological diagnosis from three major dimensions. First， at the data input level， it focuses on the characterization and preprocessing of multimodal data， including dermoscopic images， ultrasound images， and histopathological slides. Second， at the algorithmic model level， it explores ensemble learning frameworks， multimodal data fusion strategies， multicenter collaborative training approaches， and interpretable model construction. Finally， at the task recognition level， it evaluates the performance of DL models in benign skin disease screening， malignant skin lesion differentiation， and binary as well as multiclass classification tasks. By comprehensively reviewing advancements in DL-based skin disease diagnostic models from multiple perspectives， this paper aims to provide valuable insights for the further optimization and clinical translation of intelligent diagnostic systems.

Deubiquitinating enzymes are important hydrolases in the protein ubiquitin system， which regulate the dynamic balance of protein ubiquitination and deubiquitination by reversing ubiquitination， and play important roles in biological processes such as regulating protein stability， cell signal transduction， and cell apoptosis. As a member of the deubiquitinating enzyme family， ovarian tumor domain-containing protease （OTU） domain ubiquitin aldehyde binding 2 （OTUB2） can play a key role in DNA damage， signal transduction， cell metabolism， and tumorigenesis by regulating the ubiquitination levels of various substrates. OTUB2 can be abnormally expressed in various cancers， and as a regulatory factor， it significantly affects the growth， development， proliferation， migration， and invasion of cancer cells， and is closely related to the prognosis of the patients， potentially becoming a novel cancer therapeutic target and prognostic biomarker. Currently， there are many studies on the molecular mechanisms of OTUB2， but there are few review reports focusing on its role in cancer. Based on the research results about OTUB2 at home and abroad， this article reviewed the biological function of OTUB2， related signaling pathways， and its latest progress in cancer research， analyzed the application of OTUB2 in cancer treatment and prognosis evaluation， and provided new ideas for future research on OTUB2.