Objective To discuss the effect of silencing G protein-coupled receptor 139 （GPR139） gene on the proliferation， apoptosis and autophagy of breast cancer cells， and to clarify its possible mechanism. Methods The expression of GPR139 mRNA and protein in breast cancer tissue and normal tissue was downloaded from The Cancer Genome Atlas （TCGA） and Human Protein Atlas （HPA） databases； the GPR139 short hairpin RNA （shRNA） was transfected into the human breast cancer MCF-7 and MDA-MB-231 cell lines by lentiviral interference technology， and the cells were divided into sh-NC group （infected with negative control lentivirus）， sh-GPR139 group （infected with GPR139 shRNA interference lentivirus）， and sh-NC+F05 group （after infected with negative control lentivirus， GPR139 antagonist NCRW0005-F05 was added）； methylthiazolyldiphenyl-tetrazolium bromide（MTT） method was used to detect the proliferation activities of the cells in various groups； Western blotting and real-time fluorescence quantitative PCR （RT-qPCR） methods were used to detect the expression levels of proliferation markers Ki-67， Cyclin D1， Cyclin E1， and P21 mRNA and proteins in the cells in various groups； flow cytometry was used to detect the apoptotic rates of the breast cancer cells in various groups； Western blotting method was used to detect the expression levels of adenosine monophosphate-activated protein kinase （AMPK）， phosphorylated AMPK， Unc-51 like autophagy activating kinase 1 （ULK1）， phosphorylated ULK1， microtubule-associated protein 1 light chain 3A/B （LC3A/B）， and P62 proteins in the cells in various groups； immunofluorescence method was used to detected the expression level of LC3A/B protein in the breast cancer cells in various groups. Results The TCGA and HPA database results showed that the expression level of GPR139 mRNA in human breast cancer tissue was significantly higher than that in normal tissue （P<0.001）， and the expression level of GPR139 protein was significantly higher than that in normal tissue. The MTT assay results showed that compared with sh-NC group， the proliferation activities of the breast cancer cells in sh-GPR139 group and sh-NC+F05 group were significantly decreased （P<0.01）. The Western blotting results showed that compared with sh-NC group， the expression levels of GPR139， Ki67， Cyclin D1 and Cyclin E1 proteins in the breast cancer cells in sh-GPR139 group and sh-NC+F05 group were decreased （P<0.05 or P<0.01）， and the expression level of P21 protein was increased （P<0.01）. The RT-qPCR results showed that compared with sh-NC group， the expression levels of GPR139， Ki67， Cyclin D1 and Cyclin E1 mRNA in the breast cancer cells in sh-GPR139 group and sh-NC+F05 group were decreased （P<0.05 or P<0.01）， and the expression level of P21 mRNA was increased （P<0.01）. The flow cytometry results showed that compared with sh-NC group， the apoptotic rates of the breast cancer cells in sh-GPR139 group and sh-NC+F05 group were significantly increased （P<0.01）. The immunofluorescence results showed that in MCF-7 cells and MDA-MB-231 cells， compared with sh-NC group， the expression levels of LC3A/B protein in the cells in sh-GPR139 group and sh-NC+F05 group were significantly increased （P<0.01）. The Western blotting results showed that compared with sh-NC group， the expression levels of LC3A/B， p-AMPK and p-ULK1 proteins in sh-GPR139 group and sh-NC+F05 group were significantly increased （P<0.01）， and the expression level of P62 protein was significantly decreased （P<0.05 or P<0.01）. Conclusion Silencing GPR139 gene can induce autophagy and apoptosis of breast cancer cells and inhibit cell proliferation， and its mechanism may be related to the increased phosphorylation levels of AMPK and ULK1 in the cells.

Objective To discuss the functional domains of the human RNA helicase DHX37 gene predicted by online websites and software， to construct its different functional domain deletion vectors， and to verify the correctness of the constructed vectors 27. Methods Using the human DEAH-box helicase 37 （hDHX37） plasmid as a template， based on homologous recombination cloning technology， SnapGene V 6.0.2 software was used to design functional domain-specific primers for the truncations ATP binding domain （ATP binding）， C-terminal domain （CTD）， HA2 subunit （HA2）， and Oligos/Accharide binding fold domain （O/A binding fold）； prime STAR GXL polymerase， a high-fidelity DNA polymerase， was used to perform PCR amplification of the truncated target fragments ATP binding， c-terminal-1， c-terminal-2， HA2-1， HA2-2， and O/A binding； a recombination cloning kit was used to directionally clone the truncated fragments into the double-digested pCMV-MCS-3×HA-Neo empty vector； after transforming the recombinant vectors into E.coli DH5α competent cells， the correctness of the functional domain vectors was verified by restriction enzyme Hind Ⅲ digestion， agarose nucleic acid gel electrophoresis， and DNA sequencing. Results The lengths of the target fragments amplified by PCR， ATP binding （2 148 bp）， c-terminal-1 （1 326 bp）， c-terminal-2 （1 269 bp）， HA2-1 （2 205 bp）， HA2-2 （894 bp）， and O/A binding fold （2 583 bp）， were consistent with the design； Hind Ⅲ single enzyme digestion of the mutant plasmids ΔATP binding， Δc-terminal， ΔHA2， and ΔO/A binding fold yielded linearized DNA fragments with lengths of 8 939， 7 589， 8 036， 8 540， and 8 027 bp， respectively， and the product band sizes all matched the design. The DNA sequencing Blast comparative analysis results showed that ΔATP binding lacked the ATP binding domain （1-442）， Δc-terminal lacked the c-terminal domain （443-735）， ΔHA2 lacked the HA2 domain （736-861）， and ΔO/A binding fold lacked the O/A binding fold domain （862-1 158）， with deletions of 1 326， 879， 375， and 891 bp， respectively； the positions and lengths of the deleted domains in each mutant plasmid were completely consistent with the design. Conclusion The four different functional domain vectors of human DHX37 gene， ΔATP binding， Δc-terminal， ΔHA2， and ΔO/A binding fold， are successfully constructed.

Objective To discuss the effect of pyraclostrobin on ferroptosis in the spermatocytes GC-2 of the mice， and to clarify whether pyraclostrobin can cause male reproductive toxicity. Methods The mouse spermatocytes GC-2 were divided into control group， low dose of pyraclostrobin group （0.8 μmol·L^-1 pyraclostrobin）， medium dose of pyraclostrobin group （1.6 μmol·L^-1 pyraclostrobin） and high dose of pyraclostrobin group （2.4 μmol·L^-1 pyraclostrobin）. After being treated with pyraclostrobin for 24 h，methylthiazolyldiphenyl-tetrazolium bromide （MTT） method was used to detect the proliferation activities of the GC-2 cells after treated with different doses of pyraclostrobin； mitochondrial membrane potential detection kit JC-1 was used to detect the mitochondrial membrane potential in the GC-2 cells in various groups； superoxide dismutase （SOD） activity detection kit was used to detect the activities of SOD in the GC-2 cells in various groups； malondialdehyde （MDA） assay kit was used to determine the levels of MDA in the GC-2 cells in various groups； reduced glutathione （GSH） and oxidized glutathione （GSSG） detection kits were used to detect the levels of GSH and GSSG in the GC-2 cells， and the ratios of GSH/GSSG in various groups were calculated； Western blotting method was used to detect the expression levels of heme oxygenase-1 （HO-1）， ferritin heavy chain 1 （FTH1） and glutathione peroxidase 4 （GPX4） in the GC-2 cells in various groups. Results Compared with control group， the proliferation activities of the GC-2 cells in various pyaclostrobin dose ≥1.0 μg·L^-1 groups were decreased （P<0.01）. The low， medium， and high doses of pyraclostrobin were 0.8， 1.6， and 2.4 μmol·L^-1. The results of JC-1 method showed that compared with control group， the mitochondrial membrane potential of the GC-2 cells in medium， and high doses of pyraclostrobin groups were decreased. Compared with control group， the SOD activity in the GC-2 the cells in low， medium， and high doses of pyraclostrobin groups were decreased （P<0.01）， and the MDA levels in the GC-2 cells were increased （P<0.01）. Compared with control group， the GSH levels in the GC-2 cells in low， medium， and high doses of pyraclostrobin groups were significantly decreased （P<0.01）， the GSSG levels were increased （P<0.01）， and the GSH/GSSG ratios were decreased （P<0.01）. Compared with control group， the expression levels of HO-1 protein in the GC-2 cells in low， medium， and high doses of pyraclostrobin groups were significantly increased （P<0.05 or P<0.01）， while the expression levels of FTH1 and GPX4 proteins were significantly decreased （P<0.05 or P<0.01）. Conclusion Pyraclostrobin can induce the ferroptosis of the spermatocytes GC-2.

Objective To discuss the effect of iprodione （Ipr） on ferroptosis in mouse spermatocyte GC-2 cells， and to clarify its possible mechanism. Methods The MTT method was used to detect the viability of GC-2 cells after treatment with 0， 0.001， 0.010， 0.100， 1.000， 10.000， and 100.000 μmol·L^-1 Ipr for 24 h. Additional GC-2 cells were taken and divided into blank control group， 1.0 μmol·L^-1 Ipr group， 2.5 μmol·L^-1 Ipr group， and 5.0 μmol·L^-1 Ipr group （treated with Ipr solutions at final concentrations of 0， 1.0， 2.5， and 5.0 μmol·L^-1， respectively， for 24 h）. Fluorescence microscope was used to observe the intracellular reactive oxygen species （ROS） generation and mitochondrial membrane potential changes； Kit methods were used to detect the superoxide dismutase （SOD）activity， malondialdehyde （MDA） level， and the reduced glutathione （GSH）/oxidized glutathione （GSSG） ratio in the GC-2 cells in various groups； Western blotting method was used to detect the expression levels of ferroptosis-related proteins in the GC-2 cells in various groups after Ipr exposure； immunofluorescence method was used to detect the fluorescence intensity of heme oxygenase-1 （HO-1） protein in the GC-2 cells in various groups. Results The MTT assay results showed that compared with 0 μmol·L^-1 Ipr group， the viabilities of the GC-2 cells in 1.000， 10.000， and 100.000 μmol·L^-1 Ipr groups were significantly decreased （P<0.01）； thus， 1.0， 2.5， and 5.0 μmol·L^-1 Ipr were selected for subsequent experiments on GC-2 cells. The fluorescence analysis results showed that compared with blank control group， the ROS generation in the GC-2 cells in 2.5 and 5.0 μmol·L?1 Ipr groups was increased and the mitochondrial membrane potentials were decreased； there were no significant changes in the MDA level， SOD activity， and GSH/GSSG ratio in the GC-2 cells in 1.0 μmol·L^-1 Ipr group （P>0.05）； compared with blank control group， the MDA levels in the GC-2 cells in 2.5 and 5.0 μmol·L^-1 Ipr groups were significantly increased （P<0.05 or P<0.01）， and the SOD activities and GSH/GSSG ratios were significantly decreased （P<0.01）. The Western blotting results showed that compared with blank control group， after 24 h of treatment， the expression levels of glutathione peroxidase 4 （GPX4） and ferritin heavy chain 1 （FTH1） proteins in the GC-2 cells in 1.0 μmol·L^-1 Ipr group showed no significant changes （P>0.05）； compared with blank control group， after 24 h of treatment， the expression levels of GPX4 and FTH1 proteins in the GC-2 cells in 2.5 and 5.0 μmol·L?1 Ipr groups were significantly decreased （P<0.05 or P<0.01）； compared with blank control group， after 24 h of treatment， the expression levels of HO-1 protein in the GC-2 cells in 1.0， 2.5， and 5.0 μmol·L^-1 Ipr groups were significantly increased （P<0.01）. The immunofluorescence results showed that compared with blank control group， the fluorescence intensities of HO-1 protein in the GC-2 cells in 1.0， 2.5， and 5.0 μmol·L^-1 Ipr groups were significantly enhanced. Conclusion Ipr induces ferroptosis in mouse spermatocyte GC-2 cells， and its mechanism may be related to the up-regulation of HO-1 protein expression and the down-regulation of GPX4 and FTH1 protein expressions.

Objective To discuss the improvement effect of overexpressing kainate receptor subunit （GluK） 2 on the spatial learning and memory abilities of juvenile mice exposed to sevoflurane （Sevo）， and to clarify its possible molecular mechanism. Methods A total of 44 C57BL/6J neonatal mice were randomly divided into control group， Sevo group， Sevo+OE-NC group （virus empty vector group）， and Sevo+OE-GRIK2 group （GRIK2 overexpression group）. There were 3 mice in each group for molecular experiments and 8 mice in each group for behavioral experiments. Morris water maze test was used to detect the escape latency， the residence time in the target quadrant， and the number of platform crossings of the juvenile mice in various groups. The Sevo anesthesia model was established in the mice on postnatal day 6 （P6）； immunofluorescence method was used to observe the expression of GluK2 protein and the virus transfection situation in hippocampus tissue of the juvenile mice in various groups； Western blotting method was used to detect the expression levels of sodium-potassium-chloride cotransporter 1 （NKCC1）， potassium-chloride cotransporter 2 （KCC2）， and GluK2 proteins in hippocampus tissue of the juvenile mice in various groups. Results The Morris water maze test results showed that on days 3， 4， and 5 of training， compared with control group， the escape latency of the young mice in Sevo group was significantly prolonged （P<0.05 or P<0.01）. The Morris water maze test results showed that on days 4 and 5 of training， compared with Sevo group， the escape latency of the young mice in Sevo+OE-GRIK2 group was significantly shortened （P<0.05 or P<0.01）； compared with Sevo+OE-NC group， the escape latency of the young mice in Sevo+OE-GRIK2 group was significantly shortened （P<0.05 or P<0.01）. The Morris water maze test results showed that compared with control group， the residence time in the target quadrant of the young mice in Sevo group was decreased （P<0.05）； compared with Sevo group， the residence time in the target quadrant of the young mice in Sevo+OE-GRIK2 group was increased （P<0.01）； compared with Sevo+OE-NC group， the residence time in the target quadrant of the young mice in Sevo+OE-GRIK2 group was increased （P<0.01）. The Morris water maze test results showed that compared with control group， the number of platform crossings of the young mice in Sevo group was decreased （P<0.001）； compared with Sevo group， the number of platform crossings of the young mice in Sevo+OE-GRIK2 group was increased （P<0.001）； compared with Sevo+OE-NC group， the number of platform crossings of the young mice in Sevo+OE-GRIK2 group was increased （P<0.001）. The immunofluorescence results showed that compared with control group， the expression level of GluK2 protein in hippocampus tissue of the young mice in Sevo group was decreased （P<0.05）； compared with Sevo group， the expression level of GluK2 protein in hippocampus tissue of the young mice in Sevo+OE-GRIK2 group was increased （P<0.01）； compared with Sevo+OE-NC group， the expression level of GluK2 protein in hippocampus tissue of the young mice in Sevo+OE-GRIK2 group was increased （P<0.01）. The Western blotting results showed that compared with control group， the expression levels of KCC2 and GluK2 proteins in hippocampus tissue of the young mice in Sevo group were decreased （P<0.05 or P<0.01）， and the NKCC1/KCC2 ratio was increased （P<0.05）； compared with Sevo group， the expression levels of KCC2 and GluK2 proteins in hippocampus tissue of the young mice in Sevo+OE-GRIK2 group were increased （P<0.001）， and the NKCC1/KCC2 ratio was decreased （P<0.05）； compared with Sevo+OE-NC group， the expression levels of KCC2 and GluK2 proteins in hippocampus tissue of the young mice in Sevo+OE-GRIK2 group were increased （P<0.001）， and the NKCC1/KCC2 ratio was decreased （P<0.05）. Conclusion Overexpression of GRIK2 upregulates the expression levels of GluK2 and KCC2 proteins in hippocampus tissue of Sevo-exposed juvenile mice and improves the spatial learning and memory abilities of juvenile mice， and its mechanism may be related to reducing the NKCC1/KCC2 ratio in hippocampus tissue.

Objective To explore the ameliorative effect of puerarin （Pue） on non-alcoholic fatty liver disease （NAFLD） induced by high-fat diet in the mice， and to clatrify its possible mechanism. Method A total of 36 C57BL/6J mice were randomly divided into control group， model group， low dose of Pue group （20.0 mg·kg^-1）， medium dose of Pue group （40.0 mg·kg^-1）， high dose of Pue group （80.0 mg·kg^-1）， and positive drug group ［atorvastatin calcium （AT） group］ （10.0 mg·kg^-1 AT）， with 6 mice in each group. Except for control group， in which the mice were given normal diet， all of the mice in the other groups were fed with high-fat diet to establish the NAFLD models. The body weights of the mice in various groups were measured， and the liver indexes was calculated； the pathomorphology of liver tissue of the mice in various groups was observed by HE staining method. The total cholesterol （TC） and triglyceride （TG） levels in serum and liver tissue， alanine aminotransferase （ALT） and aspartate aminotransferase （AST） levels in serum， the malondialdehyde （MDA） levels， superoxide dismutase （SOD） and glutathione peroxidase （GSH-Px） activities in liver tissue， as well as interleukin-6 （IL-6）， interleukin-1β （IL-1β）， and tumor necrosis factor-α （TNF-α） levels in liver tissue of the mice in various groups were detected by commercial biochemical assay kits. Within the framework of network pharmacology， the potential therapeutic targets of Pue were predicted by integrating multiple databases， including PubChem and Swiss Target Prediction， and other publicly available resources. These targets were cross-referenced with NAFLD-associated disease targets retrieved from GeneCards and other platforms， enabling the construction of a high-confidence protein-protein interaction （PPI） network. Subsequently， Gene Ontology （GO） function and Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathway enrichment analyses were performed to elucidate the therapeutic mechanisms of Pue in NAFLD intervention； Western blotting method was used to detect the protein expression levels of Kelch-like ECH-associated protein 1 （Keap1）， nuclear factor erythroid 2-related factor 2 （Nrf2）， heme oxygenase-1 （HO-1）， and quinone oxidoreductase 1 （NQO1） in liver tissue of the mice in various groups. Additionally， 30 mice were randomly divided into control group，?model group， Pue group （80.0 mg·kg^-1 Pue gavage）， model+Nrf2 inhibitor ML385 group （30.0 mg·kg^-1 ML385 intraperitoneal administration）， and Pue+ML385 group?（80.0 mg·kg^-1 Pue gavage， 30 mg·kg^-1 ML385 intraperitoneal administration）. After treatment， the oxidative stress markers， inflammatory cytokines， and protein expression levels metioned above were detected. Results Compared with control group， the body weight and liver index of the mice in model group were significantly increased （P<0.01）； compared with model group， the body weight and liver index of the mice in low， medium， and high doses of Pue groups and AT group were significantly decreased （P<0.05 or P<0.01）. The HE staining results showed compared with control group there was significant lipid accumulation and pathological damage in liver tissue of the mice in model group； compared with model group， the accumulation of lipid droplets and pathological damage in liver tissue of the mice in low， medium， and high doses of Pue groups and AT group were improved to varying degrees. Compared with control group， the levels of TC， TG， IL-6， IL-1β， TNF-α， and MDA in liver tissue of the mice in the model group， as well as the serum TC， TG， ALT， and AST levels were significantly increased （P<0.01）， while the activities of SOD and GSH-Px in liver tissue were significantly decreased （P<0.01）； compared with model group， the levels of TC， TG， IL-6， IL-1β， TNF-α， and MDA in liver tissue， as well as the levels of TC， TG， ALT， and AST in serum of the mice in low， medium， and high doses of Pue groups and AT group were significantly decreased （P<0.05 or P<0.01）， while the activities of SOD and GSH-Px in liver tissue were significantly increased （P<0.05 or P<0.01）. The Network pharmacology analysis results identified the core targets of Pue in treating NAFLD， including Nrf2， TNF， and IL-6， and predicted that Pue may exert its effects through regulating oxidative stress， inflammation， and apoptosis pathways. The Western blotting results showed that compared with control group， the expression level of Keap1 protein in liver tissue of the mice in model group was increased （P<0.01）， while the expression levels of Nrf2， HO-1， and NQO1 proteins were decreased （P<0.01）； compared with model group， the expression levels of Keap1 protein in liver tissue of the mice in low， medium， and high doses of Pue groups and AT group were decreased （P<0.05 or P<0.01）， while the expression levels of Nrf2， HO-1， and NQO1 proteins were increased （P<0.05 or P<0.01）. After intervention with Nrf2 inhibitor ML385， compared with Pue group， the levels of IL-6， IL-1β， TNF-α， and MDA in liver tissue of the mice in Pue+ML385 group were significantly increased （P<0.05）， while the activities of SOD and GSH-Px were significantly decreased （P<0.05 or P<0.01）， the expression level of Keap1 protein in liver tissue was increased （P<0.01）， and the expression levels of Nrf2， HO-1， and NQO1 proteins were decreased （P<0.01）. Conclusion Pue can improve the lipid metabolism and liver function in the NAFLD mice， alleviate the liver damage， enhance the liver tissue antioxidant capacity， and reduce the release of inflammatory factors. Its mechanism may be related to the activation of the Keap1/Nrf2/HO-1 signaling pathway.

Objective To discuss the active components of Xinjiang characteristic plant Medicinal Mulberry Leaves against acute kidney injury （AKI） and to clarify the mechanism through ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry （UPLC-Q-TOF/MS） technology， network pharmacology， and animal experimental verification. Methods UPLC-Q-TOF/MS technology was used to analyze the chemical components in 70% ethanol percolation extract of Medicinal Mulberry Leaves and in serum of the rats at 0 and 60 min after intragastric administration； the SMILES numbers of the blood-entering active components of Medicinal Mulberry Leaves were obtained from PubMed， and then their target genes were obtained from the SIB database； meanwhile， using “acute kidney injury” as the keyword， the AKI-related targets were retrieved from databases such as the Human Gene database （GeneCards）， the Disease Gene Network database （DisGENET）， and the Online Mendelian Inheritance in Man （OMIM） disease database； the two were compared and intersected to discover the gene targets of Medicinal Mulberry Leaves for treating AKI. A total of 70 SD rats were randomly divided into blank group （10 rats） and model establishment group （60 rats）； the rats in model establishment group were intraperitoneally injected with gentamicin to establish the model for 7 d， the rats in blank group were injected with same amount of normal saline； after modeling， the serum creatinine （CRE） and urea nitrogen （BUN） levels of the rats were measured to determine whether the model was successfully prepared. A total of 60 successfully modeled rats were randomly divided into model group， low dose of Medicinal Mulberry Leaves group （120 mg·kg^-1 Medicinal Mulberry Leaves）， medium dose of Medicinal Mulberry Leaves group （300 mg·kg^-1 Medicinal Mulberry Leaves）， high dose of Medicinal Mulberry Leaves group （750 mg·kg^-1 Medicinal Mulberry Leaves）， Medicinal Mulberry Leaves enrichment substance group （750 mg·kg^-1 Medicinal Mulberry Leaves）， and positive drug group （15 mg·kg^-1 verapamil）； after treated for 14 d， the 24 h urine of the rats in various groups was collected， and the blood and kidney tissue of the rats in various groups were collected for later detection. Hematoxylin-eosin （HE） staining was used to detect the pathomorphology of kidney tissue of the rats in various groups； colorimetric method was used to detect the levels of urine protein （UP）， uric acid （UA）， β2-microglobulin （β2-MG）， albumin （ALB）， 10 kDa interferon γ-induced protein （IP-10）， kidney injury molecule-1 （KIM-1）， and neutrophil gelatinase-associated lipocalin （NGAL） in urine of the rats in various groups； microplate method was used to detect the serum creatinine （CRE） level of the rats in various groups； urease method was used to detect the serum urea nitrogen （BUN） level of the rats in various groups； ELISA method was used to detect the serum levels of interleukin-6 （IL-6） and tumor necrosis factor-α （TNF-α） of the rats in various groups and the activities of superoxide dismutase （SOD） and nitric oxide synthase （NOS） and the levels of malondialdehyde （MDA）， reduced glutathione （GSH）， and glutathione peroxidase （GSH-Px） in kidney tissue of the rats in various groups； Western blotting method was used to detect the expression levels of apoptosis and phosphatidylinositol 3-kinase （PI3K）/protein kinase B （AKT） signaling pathway-related proteins in kidney tissue of the rats in various groups. Results The UPLC-Q-TOF/MS results showed that a total of 159 chemical components were identified， and 27 blood-entering components were screened out， including 8 prototype products and 19 metabolites， mainly flavonoids， benzofurans， and organic acids. The network pharmacology results showed that AKT， B-cell lymphoma-2 （Bcl-2）， and cysteinyl aspartate specific proteinase-3 （Caspase-3） were the key targets of Medicinal Mulberry Leaves for treating AKI， and the mechanism mainly involved multi-pathway linkage and integrated regulation including the PI3K/AKT signaling pathway. Compared with model group， the levels of UP and NGAL in urine of the rats in low， medium， and high doses of Medicinal Mulberry Leaves groups were significantly decreased （P<0.01）； compared with model group， the levels of KIM-1 and IP-10 in urine of the rats in medium and high doses of Medicinal Mulberry Leaves groups were significantly decreased （P<0.01）. Compared with model group， the levels of BUN and TNF-α in blood of the rats in low， medium， and high doses of Medicinal Mulberry Leaves groups were significantly decreased （P<0.05 or P<0.01）. Compared with model group， the levels of GSH and GSH-Px in kidney tissue of the rats in medium and high doses of Medicinal Mulberry Leaves groups were significantly increased （P<0.01）； compared with model group， the level of MDA and the activity of NOS in kidney tissue of the rats in medium and high doses of Medicinal Mulberry Leaves groups were significantly decreased （P<0.01）. The Western blotting results showed that compared with model group， the expression level of PI3K protein in kidney tissue of the rats in high dose of Medicinal Mulberry Leaves group was significantly decreased （P<0.01）； compared with model group， the expression levels of AKT protein in kidney tissue of the rats in low， medium， and high doses of Medicinal Mulberry Leaves groups were decreased （P<0.05 or P<0.01）； compared with model group， the level of Bcl-2 in kidney tissue of the rats in high dose of Medicinal Mulberry Leaves group was significantly increased （P<0.05）， and the levels of Caspase-3 and Bcl-2-associated X protein （Bax） were significantly decreased （P<0.05 or P<0.01）. Conclusion Medicinal Mulberry Leaves are identified to contain a total of 159 chemical components， including 27 blood-entering components， mainly flavonoids； Medicinal Mulberry Leaves have an ameliorative effect on gentamicin-induced AKI in rats， and the mechanism may be related to the inhibition of PI3K/AKT and apoptosis signaling pathways by Medicinal Mulberry Leaves.

Objective To discuss the inhibitory effect of the bradykinin B1 receptor （B1R） antagonist ELN441958 on the growth of hepatocellular carcinoma HepG2 cells and its regulatory mechanism on the protein kinase B （Akt）/forkhead box O3a （FoxO3a） signaling pathway， and to clarify the anti-tumor effect of the B1R antagonist ELN441958 in liver cancer. Methods After the HepG2 cells were treated with different doses （0， 2.5， 5.0， 10.0， and 20.0 μmol·L^-1） of ELN441958 for 24， 48， and 72 h， 0 μmol·L^-1 ELN441958 was regared as control group. CCK-8 method was used to detect the inhibitory rate of proliferation of the cells in various groups. The HepG2 cells were divided into control group， 5.0 μmol·L^-1 ELN441958 group， 10.0 μmol·L^-1 ELN441958 group， and 15.0 μmol·L^-1 ELN441958 group； 5-ethynyl-2'-deoxyuridine （EdU） staining was used to detect the EdU positive cell rates of the HepG2 cells in various groups； flow cytometry was used to detect the apoptotic rates of the cells in various groups and the proportions of the cells at different cell cycles； immunofluorescence method was used to detect the expression level of B1R protein in the HepG2 cells in various groups； Western blotting method was used to detect the expression levels of B-cell lymphoma 2 （Bcl-2）， Bcl-2-associated X protein （Bax）， cyclin D1 （Cyclin D1）， cyclin-dependent kinase 4 （CDK4）， B1R， Akt， phosphorylated Akt （p-Akt）， FoxO3a， and phosphorylated FoxO3a （p-FoxO3a） proteins in the HepG2 cells in various groups. The HepG2 cells were divided into control group， B1R agonist des-Arg9-BK （1.0 μmol·L^-1 des-Arg9-BK） group， and BK+ELN441958 group； Western blotting method was used to detect the expression levels of Bcl-2， Bax， Cyclin D1， CDK4， Akt， p-Akt， FoxO3a， and p-FoxO3a proteins in the HepG2 cells in various groups. The HepG2 cells were divided into control group， Akt agonist SC79 group， and SC79+ELN441958 group； Western blotting method was used to detect the expression levels of Bcl-2， Bax， Cyclin D1， CDK4， Akt， p-Akt， FoxO3a， and p-FoxO3a proteins in the HepG2 cells in various groups. Results The CCK-8 assay results showed that compared with control group， when the action time was the same， the inhibitory rate of proliferation of the HepG2 cells was significantly increased with the increase of ELN441958 dose （P<0.05 or P<0.01）； when the dose of ELN441958 was the same， the inhibitory rate of proliferation of the HepG2 cells was significantly increased with the extension of action time （P<0.05 or P<0.01）. The half maximal inhibitory concentration （IC₅₀） values of ELN441958 at 24， 48， and 72 h were （21.4±1.1）， （10.5±0.3）， and （3.2±0.3） μmol·L?1， respectively. Compared with control group， the EdU positive cell rates of the HepG2 cells in 5.0， 10.0， and 15.0 μmol·L?1 ELN441958 groups were significantly decreased （P<0.05 or P<0.01）. The flow cytometry results showed that compared with control group， the apoptotic rates of the HepG2 cells in different doses of ELN441958 groups were significantly increased （P<0.05 or P<0.01）， and the percentages of the cells at G0/G1 phase were significantly increased （P<0.05 or P<0.01）. The Western blotting results showed that compared with control group， the； the expression levels of Bcl-2， CDK4， and Cyclin D1 proteins in the HepG2 cells in 5.0， 10.0 and 15.0 μmol·L^-1 ELN441958 groups were significantly decreased （P<0.01）， and the expression level of Bax protein was significantly increased （P<0.05 or P<0.01）. The immunofluorescence results showed that compared with control group， the fluorescence intensity of B1R in the HepG2 cells in 15.0 μmol·L^-1 ELN441958 group was significantly decreased （P<0.01）. The Western blotting results showed that compared with control group， the expression level of B1R， p-Akt， and p-FoxO3a proteins in the HepG2 cells in 10.0 and 15.0 μmol·L^-1 ELN441958 groups were significantly decreased （P<0.01）； the expressions p-Akt， and p-FoxO3a proteins were decreased （P<0.05 or P<0.01） compared with B1R agonist des-Arg9-BK group， the expression levels of Bcl-2， CDK4， Cyclin D1， B1R， p-Akt， and p-FoxO3a proteins in the HepG2 cells in BK+ELN441958 group were significantly decreased （P<0.01）， and the expression level of Bax protein was significantly increased （P<0.01）； compared with SC79 group， the expression levels of Bcl-2， CDK4， Cyclin D1， B1R， p-Akt， and p-FoxO3a proteins in the HepG2 cells in SC79+ELN441958 group were significantly decreased （P<0.01）， and the expression level of Bax protein was significantly increased （P<0.01）. Conclusion The B1R antagonist ELN441958 can inhibit the proliferation of HepG2 cells， and induce the apoptosis and G₀/G₁ phase cell cycle arrest of HepG2 cells， and its mechanism may be related to the regulation of Akt/FoxO3a signaling axis.

Objective To discuss the effect of inhibiting micro RNA（miR）-17-5p on scratch injury-induced proliferation of rat astrocytes and to clarify its mechanism. Methods The astrocytes were isolated and cultured primarily from rat spinal cord tissue. MiR-17-5p inhibitor and its negative control （inhibitor-NC）， and mitofusin 2 （Mfn2） interfering plasmid （si-Mfn2） and its negative control （si-NC） were transfected into rat spinal cord astrocytes. An in vitro mechanical injury-induced astrocyte proliferation model was established using the scratch method. Astrocytes were divided into control group （no treatment）， Scratch group （scratch treatment）， Scratch+inhibitor-NC group （transfected with inhibitor-NC then scratch treatment）， and Scratch+miR-17-5p inhibitor group （transfected with miR-17-5p inhibitor then scratch treatment）； and also into blank group （no treatment）， inhibitor-NC group （transfected with inhibitor-NC）， miR-17-5p inhibitor group （transfected with miR-17-5p inhibitor）， miR-17-5p inhibitor+si-NC group （co-transfected with miR-17-5p inhibitor and si-NC then scratch treatment）， and miR-17-5p inhibitor+si-Mfn2 group （co-transfected with miR-17-5p inhibitor and si-Mfn2 then scratch treatment）. Cell scratch healing assay was used to detect the scratch healing rates at different time points； real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the miR-17-5p and Mfn2 mRNA expression levels in the cells in various groups； Western blotting method was used to detect the protein expression levels of Mfn2， glial fibrillary acidic protein （GFAP）， proliferating cell nuclear antigen （PCNA）， and proliferation marker protein Ki-67 in the cells in various groups； immunofluorescence method was used to detect the expression of GFAP and Ki-67 in the cells in various groups； 5-bromo-2'-deoxyuridine （BrdU） assay was used to detect the proliferation of cells in various groups； the dual-luciferase reporter gene assay were used to verify the targeted regulatory relationship between Mfn2 and miR-17-5p. Results The cell scratch healing assay results showed that as the scratch injury time （0， 12， 24， and 48 h） increased， the scratch healing rate of spinal cord astrocytes of the rats was was increased （P<0.05） in a time-dependent manner. The RT-qPCR and Western blotting results showed that compared with Scratch 0 h group， the miR-17-5p expression level and the protein expression levels of GFAP， PCNA， and Ki-67 in astrocytes in Scratch 12， 24， and 48 h groups were increased （P<0.05）， while the Mfn2 protein expression level was decreased （P<0.05） in a time-dependent manner； compared with Control group， the GFAP and Ki-67 protein expression levels in astrocytes in Scratch group were increased （P<0.05）， while the Mfn2 protein expression level was decreased （P<0.05）； compared with Scratch group， the GFAP and Ki-67 protein expression levels in astrocytes in Scratch+miR-17-5p inhibitor group were decreased （P<0.05）， while the Mfn2 protein expression level was increased （P<0.05）； compared with blank group， the miR-17-5p expression level in astrocytes in miR-17-5p inhibitor group was decreased （P<0.05）， while the Mfn2 mRNA and protein expression levels were increased （P<0.05）； compared with Scratch+miR-17-5p inhibitor group， the Mfn2 protein expression level in astrocytes in miR-17-5p inhibitor+si-Mfn2 group was decreased （P<0.05）. The immunofluorescence assay results showed that compared with control group， the numbers of GFAP and Ki-67 co-localized astrocytes in Scratch group was increased （P<0.05）； compared with Scratch group， the number of GFAP and Ki-67 co-localized astrocytes in scratch+miR-17-5p inhibitor group was decreased （P<0.05）； compared with Scratch+miR-17-5p inhibitor group， the number of GFAP and Ki-67 co-localized astrocytes in miR-17-5p inhibitor+si-Mfn2 group was increased （P<0.05）. The BrdU assay results showed that compared with control group， the BrdU positive expression rate in the astrocytes in Scratch group was increased （P<0.05）； compared with scratch group， the BrdU positive expression rate in astrocytes in Scratch+miR-17-5p inhibitor group was decreased （P<0.05）； compared with scratch+miR-17-5p inhibitor group， the BrdU positive expression rate in astrocytes in miR-17-5p inhibitor+si-Mfn2 group was increased （P<0.05）. The dual-luciferase reporter gene assay results showed that there was a binding site between miR-17-5p and the 3'UTR region of Mfn2 mRNA， and Mfn2 is a downstream target gene of miR-17-5p. Conclusion The miR-17-5p expression level in rat spinal cord astrocytes is significantly increased after scratch injury， and cell proliferation activity is increased. Inhibition of miR-17-5p suppresses the proliferation of astrocytes induced by scratch injury by targeting and upregulating Mfn2 expression.

Objective To identify the core fatty acid metabolism-related genes （FAMRGs） in idiopathic pulmonary fibrosis （IPF） tissue using bioinformatics methods， and to experimentally verify whether these genes are significantly differentially expressed in IPF tissue and normal lung tissue. Methods Two IPF gene chip datasets， including the GSE47460 dataset and the GSE150910 dataset， were downloaded from the Gene Expression Omnibus （GEO） database； FAMRGs were screened from the Human Gene Comprehensive Database （GeneCards） database； the GEO2R tool was used to screen the differentially expressed genes （DEGs） between IPF tissue and normal lung tissue in GSE47460 dataset； Venn diagram was drawn between the DEGs and FAMRGs to obtain the differentially expressed FAMRGs. Xiantao Academic Tool was used to perform functional enrichment analysis on the above genes； the STRING database was used for analysis and Cytoscape software was used to establish a protein-protein interaction （PPI） network and screen the key FAMRGs； the GSE150910 dataset was used to verify the expression levels of the above genes， and receiver operating characteristic （ROC） curves were plotted to screen the core FAMRGs. The CIBERSORT database was used to analyze the relationship between the core FAMRGs and immune cell infiltration in IPF tissue. Eighteen healthy male SD rats were selected and randomly divided into control group， IPF group， and pirfenidone group， with 6 rats in each group. The rats in IPF group and pirfenidone group were intratracheally injected with bleomycin to establish the pulmonary fibrosis model， and the rats in control group were intratracheally injected with an equal volume of normal saline. After successful modeling， the rats in control group and IPF group were given 10 mL·kg^-1·d^-1 normal saline by gavage， and the rats in pirfenidone group were given 10 mL·kg^-1·d^-1 pirfenidone suspension by gavage； after 14 d of drug intervention， the rats were sacrificed and lung tissue was taken. Masson staining was used to observe the degree of fibrosis in lung tissue of the rats in the three groups； Western blotting method was used to detect the expression of FAMRGs proteins in lung tissue of the rats in three groups. Results A total of 182 differentially expressed FAMRGs were screened. The Gene Ontology （GO） analysis results showed that the above FAMRGs were mainly involved in functions such as hormone regulation， response to peptide substances， signaling receptor agonist activity， and cytokine activity. The Kyoto Encyclopedia of Genes and Genomes （KEGG） analysis results showed that the above FAMRGs were mainly enriched in signaling pathways such as interleukin-17 （IL-17） signaling pathway， advanced glycation end products-receptor for advanced glycation end products （AGE-RAGE） signaling pathway， and hypoxia-inducible factor-1（HIF-1） signaling pathway. Ten key FAMRGs were identified from the PPI network. Through validation with the GSE150910 dataset， matrix metalloproteinase 3 （MMP3）， secreted phosphoprotein 1 （SPP1）， and insulin-like growth factor 1 （IGF1） were determined as the core FAMRGs in IPF tissue， and their expressions were significantly higher compared with normal lung tissue （P<0.001）. The immune infiltration analysis results showed that in IPF tissue， the expressions of plasma cells， regulatory T cells， M0 macrophages， and resting mast cells were significantly up-regulated （P<0.05）， while the expressions of resting CD4 memory T cells， resting natural killer （NK） cells， monocytes， eosinophils， and neutrophils were significantly down-regulated （P<0.05）； the Spearman correlation analysis results showed that in IPF tissue， the expression level of SPP1 was positively correlated with the expression level of M0 macrophages （P<0.001）， and negatively correlated with the expression level of monocytes （P<0.001）. The Masson staining results showed that compared with control group， the collagen fiber deposition in lung tissue of the rats in IPF group was significantly increased； compared with IPF group， collagen fiber deposition in lung tissue of the rats in pirfenidone group was significantly decreased. The Western blotting results showed that compared with control group， the expression levels of MMP3， SPP1， and IGF1 proteins in lung tissue of the rats in IPF group were significantly increased （P<0.05）； compared with IPF group， the expression levels of MMP3， SPP1， and IGF1 proteins in lung tissue of the rats in pirfenidone group were significantly decreased （P<0.05）. Conclusion MMP3， SPP1， and IGF1 are the FAMRGs in IPF tissue and can serve as potential targets for future IPF treatment.

Objective To discuss the ameliorative effect of empagliflozin （EMPA） on the doxorubicin （DOX）-induced heart injury （HI） model in the rats， and to clarify its mechanism of action. Methods Twenty-four 6-7-week-old male Wistar rats were randomly divided into control group （normal healthy rats were maintained）， HI group （the DOX-induced rat HI model was established）， and HI+EMPA group （after 6 weeks of establishing the DOX-induced rat HI model， the rats were given 10 mg·kg?1 EMPA daily by gavage for 14 consecutive days）， with 8 rats in each group. Echocardiography was used to detect the left ventricular internal diameter at end-systole （LVIDs）， left ventricular ejection fraction （LVEF）， and left ventricular fractional shortening （LVFS） of the rats in various groups； HE staining and Masson staining were used to detect the pathomorphology of myocaridium tissue and collagen fibers in myocardium tissue of the rats in various groups； terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL） method was used to analyze the apoptosis in the left ventricular myocardium cells of the rats in various groups； enzyme-linked immunosorbent assay （ELISA） was used to detect the serum levels of lactate dehydrogenase （LDH） and creatine kinase （CK） of the rats in various groups. Rat myocardial H9c2 cells were used for in vitro experiments. A DOX-induced rat cardiomyocyte injury model was established in vitro （DOX group）. Cell experiment grouping-1 was divided into control group （H9c2 cells were cultured normally without any treatment）， DOX group （0.1 μmol·L^-1 DOX was added to the H9c2 cell culture medium for 48 h to induce cardiomyocyte injury）， DOX+EMPA group （0.1 μmol·L^-1 DOX and 500 nmol·L^-1 EMPA were added to the H9c2 cell culture medium for 48 h）， and DOX+EMPA+sirtuin 1 （SIRT1） inhibitor （EX527） group （H9c2 cells were pretreated with 10 μmol·L?1 EX527 for 1 h， then 0.1 μmol·L^-1 DOX and 500 nmol·L^-1 EMPA were added for 48 h）. Cell experiment grouping-2 was divided into control group （H9c2 cells were cultured normally without any treatment）， DOX group （0.1 μmol·L^-1 DOX was added to the H9c2 cell culture medium for 48 h to induce cardiomyocyte injury）， and DOX+dynamin-related protein 1 （Drp-1） inhibitor group （H9c2 cells were pretreated with 75 μmol·L^-1 Drp-1 inhibitor Mdivi-1 for 2 h， then treated with 0.1 μmol·L^-1 DOX for 48 h）. Western blotting method was used to determine the expression levels of autophagy-related proteins microtubule-associated protein 1 light chain 3 （LC3）， autophagy key molecule yeast Atg6 homolog BECLIN1， and ubiquitin-binding protein P62， SIRT1， peroxisome proliferator-activated receptor gamma coactivator 1-alpha （PGC-1α）， and Drp-1； mitochondrial fission 1 protein （Fis-1）， and mitochondrial fission factor （MFF） proteins in the cells in various groups. Results In control group， the myocardial fibers of the rat hearts were arranged neatly， and no obvious inflammatory cell infiltration or collagen fiber deposition was seen in the interstitium； compared with control group， in HI group， the myocardial fibers of the rats were disordered， the myocardial interstitium was enlarged， and inflammatory cell infiltration and collagen fiber deposition were observed； compared with HI group， in HI+EMPA group， the myocardial fibers of the rats were more regular， no obvious inflammatory cell infiltration was seen in the interstitium， and a small amount of collagen fiber deposition was observed. The TUNEL staining results showed that compared with control group， the number of TUNEL-positive cells in the myocardium tissue of the rats in HI group was increased （P<0.05）. The echocardiography results showed that compared with control group， the LVIDs of the rats in HI group was increased （P<0.05）， and the LVEF and LVFS were decreased （P<0.05）； compared with HI group， the LVIDs of the rats in HI+EMPA group was decreased （P<0.05）， and the LVEF and LVFS were increased （P<0.05）. The ELISA results showed that compared with control group， the serum levels of LDH and CK of the rats in HI group were increased （P<0.05）； compared with HI group， the serum levels of LDH and CK of the rats in HI+EMPA group were decreased （P<0.05）. The Western blotting results showed that compared with control group， the BECLIN1 protein expression level and LC3-Ⅰ/LC3-Ⅱ ratio in the H9c2 cells in DOX group were increased （P<0.05）， the expression levels of P62， SIRT1， and PGC-1α proteins were decreased （P<0.05）， and the expression level of Drp-1 protein was increased （P<0.05）； compared with DOX group， the BECLIN1 protein expression level and LC3-Ⅰ/LC3-Ⅱ ratio in the H9c2 cells in DOX+EMPA group were decreased （P<0.05）， the expression levels of P62， SIRT1， and PGC-1α proteins were increased （P<0.05）， and the expression level of Drp-1 protein was decreased （P<0.05）； compared with DOX+EMPA group， the BECLIN1 protein expression level and LC3-Ⅰ/LC3-Ⅱ ratio in the H9c2 cells in DOX+EMPA+EX527 group were increased （P<0.05）， and the expression levels of P62 and Drp-1 proteins were decreased （P<0.05）. The Western blotting results showed that compared with control group， the expression levels of Drp-1， Fis-1， and MFF proteins in the H9c2 cells in DOX group were increased （P<0.05）； compared with DOX group， the expression levels of Fis-1 and MFF proteins in the H9c2 cells in DOX+Drp-1 inhibitor group were decreased （P<0.05）. Conclusion EEMPA can alleviate DOX-induced myocardial lesions and cardiac dysfunction in rats， and reduce cardiomyocyte apoptosis and autophagy， and its mechanism may be related to the up-regulation of SIRT1 and PGC-1α protein expressions and the down-regulation of Drp1 protein expression by EMPA.

Objective To discuss the impact of an enriched environment （EE） on ischemic stroke （IS） injury， and to preliminarily clarify the role of transcription factor EB （TFEB） protein in this process as well as the relationship between EE and inflammatory response and oxidative stress response. Methods Forty?eight SD rats were randomly divided into control group， cerebral ischemia （IS） group， IS+EE group， and IS+EE+chloroquine （CQ） group （IS+EE+CQ group）， with 12 rats in each group in experiment Ⅰ. Another 16 rats were randomly divided into IS+EE+sh-NC group and IS+EE+sh-TFEB group， with 8 rats in each group in experiment Ⅱ. Before model establishment， the rats in IS+EE+sh-TFEB group were injected with TFEB shRNA into the cerebral ventricle to silence TFEB expression in brain tissue. Except for control group， the IS model was established in the rats in the other groups using the Longa suture?occlusion method. The modified neurological severity score （mNSS） was used to evaluate the neurological function injury of the rats in various groups； triphenyltetrazolium chloride （TTC） staining was used to detect the percentages of cerebral infarction area of the rats in various groups； kits were used to detect the levels of inflammatory cytokines and oxidative stress factors of the rats in various groups； Western blotting method was used to detect the expression levels of autophagy-related proteins of the rats in various groups. Results Compared with control group， the mNSS score of the rats in IS group was increased （P<0.05）， the levels of interleukin-6 （IL-6）， interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， and malondialdehyde （MDA） in the brain tissue in ischemic penumbra region were increased （P<0.05）， the activity of superoxide dismutase （SOD） was decreased （P<0.05）， and the expression levels of TFEB and Beclin-1 proteins and the ratio of microtubule-associated protein 1 light chain 3 （LC3）-Ⅱ/LC3-Ⅰ were decreased （P<0.05）. Compared with IS group， the mNSS score and the percentage of cerebral infarction area of the rats in IS+EE group were decreased （P<0.05）， the levels of IL-6， IL-1β， TNF-α， and MDA in the brain tissue in ischemic penumbra region were decreased （P<0.05）， the activity of SOD was increased （P<0.05）， and the expression levels of TFEB and Beclin-1 proteins and the ratio of LC3-Ⅱ/LC3-Ⅰ were increased （P<0.05）. Compared with IS+EE group， the mNSS score and the percentage of cerebral infarction area of the rats in IS+EE+CQ group were increased （P<0.05）， the levels of IL-6， IL-1β， TNF-α， and MDA in the brain tissue in ischemic penumbra region were increased （P<0.05）， the activity of SOD was decreased （P<0.05）， and the expression levels of TFEB and Beclin-1 proteins and the ratio of LC3-Ⅱ/LC3-Ⅰ were decreased （P<0.05）. Compared with IS+EE+sh?NC group， the mNSS score of the rats in IS+EE+sh-TFEB group was decreased （P<0.05）， the percentage of cerebral infarction area was increased （P<0.05）， and the expression levels of TFEB and Beclin-1 proteins and the ratio of LC3-Ⅱ/LC3-Ⅰ in the brain tissue in ischemic penumbra region were decreased （P<0.05）. Conclusion EE has a significant ameliorative effect on neurological function injury in the IS rats， and its mechanism may be related to EE inducing autophagy by increasing TFEB protein expression， thereby alleviating neuroinflammation and oxidative stress in the ischemic brain region.

Objective To discuss the therapeutic effect of prunetin on the mice with myocardial infarction （MI）， and to clarify the key targets and molecular mechanism of prunetin in the treatment of MI. Method Ten mice were randomly selected from 50 c57 mice as sham operation group； the other 40 mice were used to establish the MI mouse models by ligating the left anterior descending coronary artery. The 40 successfully modeled mice were randomly divided into model group， low dose of prunetin group （5 mg·kg?1 prunetin）， high dose of prunetin group （10 mg·kg?1 prunetin）， and positive drug group （2 mg·kg?1 enalapril）， with 10 mice in each group. The drugs were administered by intraperitoneal injection once daily for 21 consecutive days. Echocardiography was used to detect the cardiac function indexes of the mice in various groups； enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of creatine kinase-MB （CK-MB） and cardiac troponin Ⅰ （cTn-Ⅰ） in serum of the mice in various groups； HE staining was used to observe the patho morphology of myocardium tissue of the mice in various groups； Masson staining was used to observe the fibrosis of myocardium tissue of the mice in various groups； immunohistochemistry and immunofluorescence methods were used to detect the expression of CD31 and the number of new blood vessels in myocardium tissue at the MI border zone of the mice in various groups. The Pubchem database was used to obtain the structural information of prunetin； the SWISS and QSAR databases were used to obtain the targets of prunetin； the The GeneCards Human Gene Database （GeneCards）， Gene Web database（Disgene） and Online Mendelian Inheritance in Man （OMIM） database were used to obtain the disease targets of MI； the targets of prunetin and the disease targets were intersected to identify common targets. The String database was used to construct the protein-protein interaction （PPI） network diagram. The top-ranked targets were selected for molecular docking with prunetin to analyze the signaling pathways of prunetin in the treatment of MI. Result The echocardiography results showed that compared with sham operation group， the left ventricular ejection fraction （LVEF） and left ventricular fractional shortening （LVFS） of the mice in model group were significantly decreased （P<0.001）； compared with model group， the LVEF and LVFS of the mice in low dose and high doses of prunetin groups and enalapril group were significantly increased （P<0.05）. The ELISA results showed that compared with sham operation group， the serum levels of CK-MB and cTn-Ⅰ of the mice in model group were increased （P<0.001）； compared with model group， the levels of CK-MB and cTn-Ⅰ in the serum of the mice in low and high doses of prunetin groups and enalapril group were significantly decreased （P<0.01）. The HE staining results showed that in model group， the morphology of myocardial cells was altered， the tissue arrangement was disordered with obvious rupture， and inflammatory cell infiltration was increased； in low dose of prunetin group， the degree of myocardial cell necrosis was alleviated； in high dose of prunetin group， the symptoms of myocardial cell swelling， disordered arrangement， and myofibril rupture were significantly relieved. The Masson staining results showed that in model group， myocardial fibrosis was obvious and the anterior ventricular wall was thinned， while in low dose of prunetin group， the myocardial fibrosis area was reduced， significantly inhibiting the development of fibrosis， and in high dose of prunetin group， the myocardial fibrosis area was significantly decreased， and the scar-like changes caused by fibrosis were alleviated. The immunohistochemistry and immunofluorescence results showed that compared with model group， the number of new blood vessels in the MI border zone myocardium tissue of the mice in high dose of prunetin treatment group was increased （P<0.05）. The network pharmacology results showed that there were 100 compound targets of prunetin， among which 68 were common targets for prunetin-MI. The key genes related to MI included epidermal growth factor receptor （EGFR）， phosphatidylinositol 3-kinase catalytic subunit alpha（PIK3CA）， and peroxisome proliferator-activated receptor gamma（PPARG）， etc. The molecular docking analysis results showed that prunetin docked well with the key targets EGFR and PIK3CA， and had better affinity with EGFR. Conclusion Prunetin can promote the angiogenesis of capillaries in the infarct border zone of the MI mouse models to alleviate ischemia-hypoxia injury， thereby alleviating cardiac dysfunction in mice after MI.

Objective To discuss the role of microRNA （miR）-378 in cardiomyocyte injury induced by myocardial ischemia reperfusion （I/R）， and to clarify its mechanism. Methods The human cardiomyocytes AC16 and human monocyte macrophages THP-1 were cultured. Firstly， miR-378 mimics and its negative control （miR-NC） were transfected into the THP-1 cells respectively.The cells were divided into control group， miR-NC group and miR-378 mimics group， the THP-1 cells in control group were cultured normally. Real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression level of miR-378 and the expression levels of M1 macrophage secreted factors ［inducible nitric oxide synthase （iNOS）， tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β） and interleukin-6 （IL-6）］ and M2 macrophage secreted factors ［arginase-1 （Arg-1）， transforming growth factor-β1 （TGF-β1）， interleukin-4 （IL-4） and interleukin-10 （IL-10）］ in the THP-1 cells after transfected； Western blotting method was used to detect the expression levels of M1 macrophage surface marker protein cluster of differentiation 86 （CD86） and M2 macrophage surface marker protein cluster of differentiation 206 （CD206）. The AC16 cells underwent hypoxia/reoxygenation （H/R） treatment to simulate I/R cardiomyocyte injury， and were co-cultured with differently treated THP-1 cells. The cells were divided into control group （upper chamber containing AC16 cells， lower chamber containing THP-1 cells）， H/R group （upper chamber containing H/R-induced AC16 cells， lower chamber containing THP-1 cells）， miR-NC+H/R group （upper chamber containing H/R-induced AC16 cells， lower chamber containing THP-1 cells transfected with miR-NC） and miR-378 mimics+H/R group （upper chamber containing H/R-induced AC16 cells， lower chamber containing THP-1 cells transfected with miR-378 mimics）. Cell counting kit-8 （CCK-8） method was used to detect the activities of the AC16 cells in various groups； kits were used to detect the levels of malondialdehyde （MDA） and reactive oxygen species （ROS） and the activity of superoxide dismutase （SOD） in the AC16 cells in various groups as well as the lactate dehydrogenase （LDH） activity in the cell supernatant. Results The RT-qPCR method and Western blotting method results showed that compared with control group and miR-NC group， the expression level of miR-378 in the THP-1 cells in miR-378 mimics group was significantly increased （P<0.05）； the expression levels of iNOS， TNF-α， IL-1β， and IL-6 mRNA and the expression level of CD86 protein in the THP-1 cells were significantly decreased （P<0.05）； the expression levels of Arg-1， TGF-β1， IL-4， and IL-10 mRNA and the expression level of CD206 protein in the THP-1 cells were significantly increased （P<0.05）. The CCK-8 method and kit method results showed that compared with control group， the activity of the AC16 cells in H/R group was significantly decreased （P<0.05）， the levels of MDA and ROS in the cells were significantly increased （P<0.05）， the activity of SOD was significantly decreased （P<0.05）， and the LDH activity in the cell supernatant was significantly increased （P<0.05）. Compared with H/R group， the activity of the AC16 cells in miR-378 mimics+H/R group was significantly increased （P<0.05）， the levels of MDA and ROS in the cells were significantly decreased （P<0.05）， the activity of SOD was significantly increased （P<0.05）， and the LDH activity in the cell supernatant was significantly decreased （P<0.05）. Conclusion miR-378 can alleviate H/R-induced cardiomyocyte injury， and its mechanism may be related to regulating macrophage polarization towards the M2 phenotype.

Objective To discuss the effects of transient receptor potential vanilloid channel 2 （TRPV2） on the proliferation， migration， and invasion of oral squamous cell carcinoma （OSCC） cells and the effect of cannabidiol （CBD） on the biological behaviors of OSCC cells through the TRPV2 channel， and to clarify its related anti-tumor mechanism. Methods The siRNA fragments were transfected into the CAL-27 cells using the liposome method， and the cells were divided into si-NC group （transfected with non-related si-NC） and si-TRPV2 group （transfected with specific siRNA silencing TRPV2 gene）. The CAL-27 cells were cultured with different doses of CBD and divided into 0， 10， 20， and 40 μmol·L^-1 CBD groups. Real-time fluorescence quantitative PCR （RT-qPCR） method and Western blotting method were used to detect the expression levels of TRPV2 mRNA and protein in the cells in various groups； cell counting kit-8 （CCK-8） method， cell scratch healing assay， and Transwell chamber assay were used to detect the proliferation activity， scratch healing rate， and number of invasion CAL-27 cells in various groups， respectively. Results The RT-qPCR method and Western blotting method results showed that the expression levels of TRPV2 mRNA and protein in the CAL-27 cells in si-NC group were higher than those in si-TRPV2 group （P<0.01）. The CCK-8 assay results showed that compared with si-NC group， the proliferation activities of the cells in si-TRPV2 group at 48 and 72 h after transfection were significantly decreased （P<0.01）. The cell scratch healing assay results showed that compared with si-NC group， the scratch healing rate of the CAL-27 cells in si-TRPV2 group at 24 h was significantly decreased （P<0.01）. The Transwell chamber assay results showed that compared with si-NC group， the number of invasion cells in si-TRPV2 group was significantly decreased （P<0.01）. The RT-qPCR method and Western blotting method results showed that compared with 0 μmol·L^-1 CBD group， the expression levels of TRPV2 mRNA and protein in the CAL-27 cells in 10， 20， and 40 μmol·L^-1 CBD groups were decreased （P<0.05 or P<0.01）. The CCK-8 assay results showed that compared with 0 μmol·L^-1 CBD group， the cell survival rates of the cells in 20 and 40 μmol·L^-1 CBD groups at 48， 72， and 96 h after drug culture were significantly decreased （P<0.01）. The cell scratch healing assay results showed that after CBD culture for 12 and 24 h， compared with 0 μmol·L^-1 CBD group， the scratch healing rates of the CAL-27 cells in 10， 20， and 40 μmol·L^-1 CBD groups were significantly decreased （P<0.01）. The Transwell chamber assay results showed that compared with 0 μmol·L^-1 CBD group， the number of invasion cells in 40 μmol·L^-1 CBD group was significantly decreased （P<0.01）. Conclusion Silencing the TRPV2 gene and CBD treatment can both inhibit the proliferation， migration， and invasion abilities of CAL-27 cells， and CBD can reduce the expression of TRPV2 gene and protein in CAL-27 cells.

Objective To discuss the inductive effect of lysophosphatidic acid （LPA） combined with 6-hydroxydopamine （6-OHDA） on the apoptosis of SH-SY5Y cells， and to clarify its related mechanism. Methods The Parkinson’s disease （PD） cell model was established by using 100 μmol·L^-1 6-OHDA； the SH-SY5Y cells， PC12 cells and N2a cells were selected and divided into control group （incomplete medium only）， 4 μmol·L^-1 LPA group， 100 μmol·L^-1 6-OHDA group， 4 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group， 10 μmol·L^-1 LPA group， and 10 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group， and treated for 24 h. Methylthiazolydiphenyl-tetrazolium bromide（MTT） method was used to detect the activities of the SH-SY5Y cells in various groups； annexin Ⅴ fluorescein isothiocyanate（Annexin Ⅴ-FITC）/propidium iodide（PI） staining and flow cytometry were used to detect the apoptotic rates of the SH-SY5Y cells in various groups； Western blotting method was used to detect the expression levels of Caspase-3 related proteins in three kinds of cells and the expression levels of B-cell lymphoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， p38 mitogen-activated protein kinase （p38 MAPK）， phosphorylated p38 MAPK （p-p38 MAPK） proteins and LPA1 receptor protein in the SH-SY5Y cells in various groups. Results The MTT assay results showed that compared with 100 μmol·L?1 6-OHDA group， the numbers and activities of the SH-SY5Y cells in 4 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group and 10 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group were significantly decreased （P<0.05 or P<0.01）. The flow cytometry results showed that compared with 100 μmol·L^-1 6-OHDA group， the apoptotic rates of the SH-SY5Y cells in 4 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group and 10 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group were increased （P<0.05 or P<0.01）. The Western blotting results showed that compared with 100 μmol·L^-1 6-OHDA group， the expression levels of Caspase-3 protein in three kinds of cells in 4 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group and 10 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group had no significant changes， and the differences were not statistically significant （P>0.05）； the expression levels of Cleaved Caspase-3 protein in the SH-SY5Y cells in 4 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group were significantly increased （P<0.01）； the expression levels of Cleaved Caspase-3 protein in the PC12 cells in 4 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group and 10 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group were significantly increased （P<0.05 or P<0.01）； the expression level of Cleaved Caspase-3 protein in the N2a cells in 4 μmol·L?1 LPA+100 μmol·L?1 6-OHDA group was increased （P<0.05）. The Western blotting results showed that compared with 100 μmol·L^-1 6-OHDA group， the expression levels of Bax and Bcl-2 proteins in the SH-SY5Y cells in 4 μmol·L^-1 LPA+100 μmol·L?1 6-OHDA group and 10 μmol·L?1 LPA+100 μmol·L?1 6-OHDA group had no significant changes， and the differences were not statistically significant （P>0.05）， but the Bax/Bcl-2 ratio was increased （P<0.05）； the expression level of p38 MAPK protein in the SH-SY5Y cells in 10 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group was increased （P<0.05）； the expression level of p-p38 MAPK protein in the SH-SY5Y cells in 4 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group was significantly increased （P<0.01）， and the p-p38 MAPK/p38 MAPK ratio was increased （P<0.05）； the expression level of LPA1 receptor protein in the SH-SY5Y cells in 10 μmol·L^-1 LPA+100 μmol·L^-1 6-OHDA group was decreased （P<0.05）. Conclusion The combined application of LPA and 6-OHDA can significantly induce the apoptosis of SH-SY5Y cells， and its mechanism may be related to the up-regulated expression levels of p38 MAPK and p-p38 MAPK proteins in the cells.

Objective To discuss the protective effect of Liangge Powder （LGS） on lipopolysaccharide （LPS）-induced acute lung injury （ALI）， and to clarify its mechanism. Methods Forty male C57BL/6 mice were randomly divided into sham operation group （SHMA group）， model group （LPS group）， low dose of LGS group （LGSL group， 8.58 g·kg?1 LGS）， and high dose of LGS group （LGSH group， 17.16 g·kg?1 LGS）， with 10 mice in each group. The mice in LPS group， LGSL group， and LGSH group were intratracheally instilled with LPS to induce the ALI model， and samples were taken 24 h after modeling. HE staining was used to observe the pathomorphology of lung tissue of the mice in various groups and to perform the pathological scores； flow cytometry was used to detect the percentages of neutrophils and epithelial cells in lung tissue of the mice in various groups； immunohistochemistry staining assay was used to detect the level of myeloperoxidase （MPO） in lung tissue of the mice in various groups； enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of related inflammatory mediators interleukin-18 （IL-18）， interleukin-1β （IL-1β）， and albumin in bronchoalveolar lavage fluid of the mice in various groups； Western blotting method was used to detect the expression levels of high mobility group box 1 （HMGB1）， NLR family pyrin domain containing 3 （NLRP3）， cysteinyl aspartate specific proteinase-1 （Caspase-1）， and gasdermin D （GSDMD） proteins in lung tissue of the mice in various groups. Results The HE staining results showed that compared with SHMA group， the pathological injury of lung tissue of the mice in LPS group was aggravated， and the pathological score was significantly increased （P<0.05）； compared with LPS group， the pathological scores of lung tissue of the mice in LGSL group and LGSH group were significantly decreased （P<0.05）. The flow cytometry results showed that compared with SHMA group， the percentage of neutrophils in lung tissue of the mice in LPS group was increased （P<0.05）， and the percentage of epithelial cells was decreased （P<0.05）； compared with LPS group， the percentages of neutrophils in lung tissue of the mice in LGSL group and LGSH group were decreased （P<0.05）， and the percentages of epithelial cells were increased （P<0.05）. The immunohistochemistry staining results showed that compared with SHMA group， the MPO level in lung tissue of the mice in LPS group was significantly increased （P<0.05）； compared with LPS group， the MPO levels in lung tissue of the mice in LGSL group and LGSH group were significantly decreased （P<0.05）. The ELISA results showed that compared with SHMA group， the levels of IL-1β， IL-18， and albumin in bronchoalveolar lavage fluid of the mice in LPS group were significantly increased （P<0.05）； compared with LPS group， the levels of IL-1β， IL-18， and albumin in bronchoalveolar lavage fluid of the mice in LGSL group and LGSH group were significantly decreased （P<0.05）. The Western blotting results showed that compared with SHMA group， the expression levels of HMGB1， NLRP3， Caspase-1， and GSDMD proteins in lung tissue of the mice in LPS group were significantly increased （P<0.01）； compared with LPS group， the expression levels of HMGB1， NLRP3， Caspase-1， and GSDMD proteins in lung tissue of the mice in LGSL group and LGSH group were significantly decreased （P<0.05）. Conclusion LGS can alleviate LPS-induced pathological lung injury and inflammatory response by reducing neutrophil infiltration and epithelial cell pyroptosis， and its mechanism may be related to the down-regulation of NLRP3， Caspase-1， and GSDMD protein expressions through the HMGB1 signaling pathway.

Objective To discuss the effect of upstream transcription factor 2 （USF2） on coagulation dysfunction in the septic rats， and to clarify its potential mechanism based on the protein tyrosine phosphatase non-receptor type 2 （PTPN2）/c-Jun N-terminal kinase （JNK）/sterol regulatory element-binding protein 2 （SREBP2） signaling pathway. Methods Fifteen healthy SD rats were randomly selected from 265 rats as control group （no ligation or puncture）； the remaining 250 rats were used to establish the sepsis models by cecal ligation and puncture （CLP）. Seventy-five successfully modeled rats were randomly divided into model group （CLP）， positive drug group （CLP+20 mg·kg?1 simvastatin）， small interfering RNA （siRNA） negative control （si-NC） group （CLP+transfection with si-NC）， si-USF2 group （CLP+transfection with USF2-siRNA）， and JNK activator group （CLP+transfection with USF2-siRNA+2 mg·kg?1 JNK activator Anisomycin）， with 15 rats in each group. An automatic hematology analyzer was used to detect the platelet （PLT） count of the rats in various groups； an automatic coagulation analyzer was used to detect the prothrombin time （PT）， activated partial thromboplastin time （APTT）， thrombin time （TT）， and the levels of D-dimer （DD） and fibrinogen （FIB） of the rats in various groups； enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of inflammatory factors including interleukin-1β （IL-1β）， interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， C-reactive protein （CRP）， and procalcitonin （PCT） in serum of the rats in various groups； kits were used to detect the superoxide dismutase （SOD） activity and the levels of malondialdehyde （MDA） and glutathione （GSH） in serum of the rats in various groups； HE staining was used to observe the pathomorphology of lung tissue and cecal tissue of the rats in various groups； real-time fluorescence quantitative PCR （RT-qPCR） and Western blotting methods were used to detect the expression levels of USF2 mRNA and protein and the expression levels of PTPN2， phosphorylated JNK （p-JNK）， JNK， and SREBP2 proteins in lung tissue and cecal tissue of the rats in various groups. Results After 12 d of modeling， the survival rate of the rats in control group was significantly higher than that in model group. Compared with control group， the expression levels of USF2 mRNA and protein in lung tissue and cecal tissue of the rats in model group， positive drug group， si-NC group， si-USF2 group， and JNK activator group were significantly increased （P<0.05）； compared with model group and si-NC group， the expression levels of USF2 mRNA and protein in lung tissue and cecal tissue of the rats in si-USF2 group and JNK activator group were significantly decreased （P<0.05）. Compared with control group， the PLT counts of the rats in model group， positive drug group， si-NC group， si-USF2 group， and JNK activator group were significantly decreased （P<0.05）； compared with model group， the PLT counts of the rats in positive drug group， si-USF2 group， and JNK activator group were significantly increased （P<0.05）； compared with si-NC group， the PLT counts of the rats in si-USF2 group and JNK activator group were significantly increased （P<0.05）； compared with si-USF2 group， the PLT count of the rats in JNK activator group was significantly decreased （P<0.05）. Compared with control group， the APTT， PT， TT and DD levels of the rats in model group， positive drug group， si-NC group， si-USF2 group， and JNK activator group were significantly increased （P<0.05）， and the FIB level was significantly decreased （P<0.05）； compared with model group， the APTT， PT， TT and DD levels of the rats in positive drug group， si-USF2 group， and JNK activator group were significantly decreased （P<0.05）， and the FIB level was significantly increased （P<0.05）； compared with si-NC group， the APTT， PT， TT and DD levels of the rats in si-USF2 group and JNK activator group were significantly decreased （P<0.05）， and the FIB level was significantly increased （P<0.05）； compared with si-USF2 group， the APTT， PT， TT and DD levels of the rats in JNK activator group were significantly increased （P<0.05）， and the FIB level was significantly decreased （P<0.05）. Compared with control group， the levels of IL-1β， IL-6， TNF-α， CRP， PCT and MDA in serum of the rats in model group， positive drug group， si-NC group， si-USF2 group， and JNK activator group were significantly increased （P<0.05）， and the SOD activity and GSH level were significantly decreased （P<0.05）； compared with model group， the levels of IL-1β， IL-6， TNF-α， CRP， PCT and MDA in serum of the rats in positive drug group， si-USF2 group， and JNK activator group were significantly decreased （P<0.05）， and the SOD activity and GSH level were significantly increased （P<0.05）； compared with si-NC group， the levels of IL-1β， IL-6， TNF-α， CRP， PCT and MDA in serum of the rats in si-USF2 group and JNK activator group were significantly decreased （P<0.05）， and the SOD activity and GSH level were significantly increased （P<0.05）； compared with si-USF2 group， the levels of IL-1β， IL-6， TNF-α， CRP， PCT and MDA in serum of the rats in JNK activator group were significantly increased （P<0.05）， and the SOD activity and GSH level were significantly decreased （P<0.05）. Compared with control group， the alveolar structure of the lung tissue of the rats in model group rats was damaged， the villi of the cecal tissue disappeared， and a large number of inflammatory cells infiltrated； compared with model group， the lung tissue alveolar damage and cecal villi damage of the rats in positive drug group， si-USF2 group， and JNK activator group were alleviated， and inflammatory cell infiltration was reduced； compared with si-NC group， the above pathological changes in the lung and cecal tissues of the rats in si-USF2 group and JNK activator group were significantly alleviated； compared with si-USF2 group， the pathological changes in the lung and cecal tissues of rats in JNK activator group were aggravated.Compared with control group， the expression level of SREBP2 protein and the p-JNK/JNK ratio in lung tissue and cecal tissue of the rats in model group， positive drug group， si-NC group， si-USF2 group， and JNK activator group were significantly increased （P<0.05）， and the expression level of PTPN2 protein was significantly decreased （P<0.05）； compared with model group and si-NC group， the expression level of SREBP2 protein and the p-JNK/JNK ratio in lung tissue and cecal tissue of the rats in positive drug group， si-USF2 group， and JNK activator group were significantly decreased （P<0.05）， and the expression level of PTPN2 protein was significantly increased （P<0.05）； compared with si-NC group， the expression level of SREBP2 protein and the p-JNK/JNK ratio in lung tissue and cecal tissue of the rats in si-USF2 group and JNK activator group were significantly decreased （P<0.05）， and the expression level of PTPN2 protein was significantly increased （P<0.05）； compared with si-USF2 group， the expression level of SREBP2 protein and the p-JNK/JNK ratio in lung tissue and cecal tissue of the rats in JNK activator group were significantly increased （P<0.05）， and the expression level of PTPN2 protein was significantly decreased （P<0.05）. Conclusion Knockdown of USF2 gene can significantly improve the pathomorphology of lung tissue and cecal tissue， alleviate coagulation dysfunction， and reduce the levels of inflammatory factors and oxidative stress in septic rats； its mechanism may be related to the regulation of PTPN2/JNK/SREBP2 signaling pathway.

Objective To discuss the induction effect of wedelolactone （WEL） on cuproptosis in the human pancreatic cancer cells （PANC-1）， and to clarify its molecular mechanism. Methods The PANC-1 cells were treated with different concentrations （0-300 μmol·L^-1） of WEL for 12， 24， and 48 h， respectively， cell counting kit-8 （CCK-8） method was used to detect the survival rates of the cells after treated with different concentrations of WEL to determine the drug concentration and action time for subsequent experiments. The human pancreatic cancer PANC-1 cells were divided into control group （0 μmol·L^-1 WEL）， 8.75 μmol·L^-1 WEL group， 17.50 μmol·L^-1 WEL group， and 35.00 μmol·L^-1 WEL group. Colony formation assay was used to detect the colony formation rates of the PANC-1 cells in various groups； 5-ethynyl-2'-deoxyuridine （EdU） staining was used to detect the EdU positive cell rates of the cells in various groups； lactate dehydrogenase （LDH） kit was used to detect the LDH release in supernatant of the cells in various groups. After treating PANC-1 cells with caspase inhibitor （Z-VAD-FMK）， cuproptosis inhibitor tetrathiomolybdate （TTM）， ferroptosis inhibitor ferrostatin-1 （Fer-1）， and necroptosis inhibitor necrostatin-1 （Nec-1） combined with 35.00 μmol·L^-1 WEL for 48 h， CCK-8 method was used to detect the cell survival rates of the PANC-1 cells after treated with different inhibitors， and to screen the death mode induced by WEL； cell copper （Cu2?） colorimetric assay kit was used to detect the intracellular Cu2? levels of the cells in various groups； transmission electron microscope was used to observe the mitochondrial ultrastructure of the PANC-1 cells in various groups； mitochondrial membrane potential assay kit （JC-1） was used to detect the mitochondrial membrane potential of the cells in various groups； immunofluorescence staining was used to detect the expression and mitochondrial co-localization of dihydrolipoamide S-acetyltransferase （DLAT） in the cells in various groups； Western blotting method was used to detect the expression levels of ferredoxin 1 （FDX1）， lipoic acid synthetase （LIAS）， DLAT， and dihydrolipoamide S-succinyltransferase （DLST） proteins in the cells in various groups. Results The CCK-8 assay results showed that compared with control group， the survival rates of the PANC-1 cells after treated with different concentrations of WEL for 12， 24， and 48 h were significantly decreased （P<0.05）， with the most significant inhibitory effect at 48 h； therefore， 0， 8.75， 17.50， and 35.00 μmol·L^-1 WEL were selected to treat the PANC-1 cells. The colony formation assay results showed that compared with control group， the colony formation rates of the PANC-1 cells in 8.75， 17.50， and 35.00 μmol·L^-1 WEL groups were decreased （P<0.01）. The EdU assay results showed that compared with control group， the EdU positive cell rates of the PANC-1 cells in 8.75， 17.50， and 35.00 μmol·L^-1 WEL groups were decreased （P<0.01）. The LDH assay results showed that compared with control group， the LDH release in supernatant of PANC-1 cells in 8.75， 17.50， and 35.00 μmol·L^-1 WEL groups was increased （P<0.01）. The cell Cu2? colorimetric assay kit results showed that compared with control group， the Cu2? levels in the PANC-1 cells in 8.75， 17.50， and 35.00 μmol·L?1 WEL groups were increased （P<0.01）. The inhibitor intervention assay results showed that compared with control group， the survival rate of the PANC-1 cells in 35.00 μmol·L?1 group was decreased（P<0.01）； compared with 35.00 μmol·L?1 WEL group， the survival rates of the PANC-1 cells in WEL+Z-VAD-FMK group and WEL+TTM group were increased （P<0.01）. The transmission electron microscope results showed that the mitochondria in the PANC-1 cells in 35.00 μmol·L^-1 WEL group exhibited membrane rupture， reduced number of cristae， and sparse arrangement. The JC-1 staining results showed that compared with control group， the mitochondrial membrane potential in the PANC-1 cells in 17.50 and 35.00 μmol·L^-1 WEL groups was significantly decreased （P<0.01）. The immunofluorescence staining results showed that compared with control group， the DLAT fluorescence intensity in the PANC-1 cells in 17.50 and 35.00 μmol·L^-1 WEL groups was significantly increased （P<0.01） and co-localized with mitochondria. The Western blotting method results showed that compared with control group， the expression level of FDX1 protein in the PANC-1 cells in 8.75 μmol·L?1 WEL group was increased （P<0.01）， the expression levels of DLAT， DLST， and FDX1 proteins in the PANC-1 cells in 17.50 and 35.00 μmol·L?1 WEL groups were significantly increased （P<0.01）， while the expression level of LIAS protein was significantly decreased （P<0.01）. Conclusion WEL can induce cuproptosis in the PANC-1 cells， and its mechanism may be related to increasing the Cu2? level and up-regulating the expression levels of key cuproptosis proteins DLAT， DLST， and FDX1 in the PANC-1 cells.

Objective To explore the association between atherogenic index of plasma （AIP） and the risk of hypertension， and to clarify the application value of AIP as a risk predictor for hypertension. Methods The subjects were derived from a prospective cohort population established using a multi-stage stratified cluster sampling method. Based on the AIP quartiles， the included subjects were divided into four groups （Q1 group， Q2 group， Q3 group， and Q4 group）. The association between AIP and the risk of hypertension was explored using the Cox regression model， and subgroup analysis and interaction tests were employed to assess gender and age differences in this association. Results A total of 2 532 subjects were included in this study （632 in Q1 group， 634 in Q2 group， 633 in Q3 group， and 633 in Q4 group）. The median follow-up time was 4.69 years， and 1 587 subjects eventually developed hypertension （the incidence rate was 62.68%）. Compared with Q1， Q2， and Q3 groups， the male proportion in Q4 group was 35.70%， and a smoking proportion was 36.90%； the body mass index（BMI） and fasting plasma glucose（FPG） levels （P<0.001）. Based on a cohort study and multiple factor Cox regression model analysis， compared with Q1 group， the risk of hypertension in Q4 groups was increased by 24% ［hazard ratio（HR）=1.24， 95% confidence interval （CI）=1.07-1.44， P=0.004］. The interaction test results and subgroup analysis showed that there was an interaction between age and gender and AIP （P<0.05）； in the female population （HR=1.42， 95%CI=1.16-1.73， P=0.001） and<60 year old population （HR=1.31， 95%CI=1.11-1.54， P=0.001）， high AIP was a risk factor for developing hypertension. Conclusion An elevated AIP is associated with a higher risk of hypertension， particularly in the female and<60 year old populations where a higher AIP is closely related to the occurrence of hypertension.

Objective To analyze the temporal evolution pattern of the disease burden of liver cancer in China from 1990 to 2021， to construct a disease burden prediction system， to simulate the evolution trend of the disease spectrum up to 2050， and to identify core pathogenic factor clusters through an attribution risk analysis model， and to provide the evidence-based support for the formulation of public health policies. Methods Based on the Global Burden of Disease Study 2021 （GBD 2021） database， the trends in the incidence， mortality， and disability-adjusted life years （DALYs） of liver cancer in China from 1990 to 2021 were systematically analyzed； the Bayesian age-period-cohort （BAPC） model was used to predict the evolution trajectory up to 2050； the main attributable risk factors were evaluated， including smoking， alcohol abuse， high body mass index （BMI）， high fasting plasma glucose， and drug use. Results From 1990 to 2021， the age-standardized incidence rate （ASIR）， age-standardized mortality rate （ASMR）， and age-standardized disability-adjusted life years rate （ASDR） of liver cancer in China showed decreasing trends， with the estimated annual percentage change （EAPC） showing decreasing trends （-0.28， -0.76， and -0.82）， respectively， and the decrease was greater in females than in males； during the same period， the absolute number of cases， deaths， and total DALYs of liver cancer continued to increase. In 2021， the main attributable factors were smoking （13.71%）， alcohol use （11.49%）， and metabolic abnormalities （high BMI was 7.40% and high fasting plasma glucose was 2.01%）； the attribution patterns were different among different age and sex groups； the impact of smoking on liver cancer deaths in males was significantly higher than that in females； the burden of alcohol abuse and drug use was concentrated in the 70-74 years old age group； the peak risk exposure of high BMI shifted forward to the 45-49 years old age group； while the risk aggregation of high fasting plasma glucose was significantly delayed to the advanced age group of 80-84 years old. The BAPC model predicted that the ASIR， ASMR， and ASDR of liver cancer in China would continue to decrease by 2050. Conclusion Although the standardized burden of liver cancer in China shows a decreasing trend， its absolute number is still increasing； the main attributable risk factors， including smoking， alcohol abuse， and metabolic abnormalities （high BMI and high fasting plasma glucose）， show differential distributions among different age and sex groups. This suggests that future prevention and control strategies for liver cancer should focus on the multi-dimensional management of behavioral and metabolic risk factors， and implement precise preventive measures combined with the sex and age differences of the liver cancer patients.

Objective To establish a three-dimensional finite element model of mandibular teeth during retraction in sliding mechanics， by simulating the application of different torque controls of anterior teeth， and to clarify the impact of different torque controls on the movement of mandibular teeth during the retraction process. Methods Using cone beam computed tomography （CBCT） data from a permanent dentition patient scheduled for mandibular first premolar extraction in orthodontic treatment， a three-dimensional finite element model of straight-wire sliding retraction of mandibular teeth was constructed via Mimics， Geomagic Wrap， and Solidworks software. In Ansys Workbench software， six conditions with different torques of -11°， -6°， -3°， +3°， +6° and+11° were applied to the mandibular anterior teeth， with a 1.5 N retraction force loaded. The initial displacement trends of each mandibular teeth in the labiolingual， mesiodistal， and vertical directions were analyzed under these conditions. Results In the buccolingual directions， different anterior torques exerted a tendency to procline and intrude the central and lateral incisors， with this tendency becoming increasingly evident as the torque increased. For other teeth in the lower arch， it induced a lingual tip and extrusion trend， with the canines and second premolars being the most affected by torque changes， followed by the first molars. The second molars were primarily influenced by the retraction force， exhibiting a displacement trend of mesiallingual rotation and were less affected by torque changes. In the mesialdistal direction， the changes in anterior torques had no significant impact on the posterior teeth. Conclusion Different anterior torques lead to extrusion and lingual tipping movements of other teeth in the mandibular arch， with this influence also depending on the position of the teeth within the arch. Teeth located more posteriorly are less affected by torque changes but are more influenced by the retraction force. When applying different torques to control anterior teeth clinically， the clinicans should pay more attention to the movements of other teeth.

Objective To investigate the potential causal associations of specific plasma lipids with nonalcoholic fatty liver disease （NAFLD） and liver enzymes， and elucidate the potential impact of specific plasma lipid on liver health. Methods Two-sample Mendelian randomization （MR） analyses were conducted to investigate potential causal relationships between 179 plasma lipids and both NAFLD and 4 liver enzymes： alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， gamma-glutamyl transferase （GGT）， and alkaline phosphatase （ALP）. The inverse-variance weighted method served as the primary analysis approach， supplemented by MR-Egger， weighted median， weighted mode， and simple mode. Sensitivity analyses including MR-Egger intercept test and MR-PRESSO Global test were conducted to ensure the reliability of the results. Multivariable MR analysis was conducted to adjust for the potential effect of body mass index （BMI） on the association between plasma lipids and NAFLD. Results The analyses revealed that phosphatidylcholine （18：0/20：5） ［odd ratio（OR）=0.988， 95% confidence interval（CI） （0.977， 0.999）， P=0.028］ exhibited a protective effect on NAFLD， while phosphatidylinositol （16：0/18：2） ［OR=1.016， 95%CI（1.000， 1.032）， P=0.046］ and phosphatidylinositol （18：0/18：2） ［OR=1.012， 95%CI （1.002， 1.022）， P=0.021］ increased NAFLD risk. After adjustment for BMI， the association between phosphatidylinositol （18：0/18：2） ［OR=1.019， 95%CI （1.007， 1.035）， P=0.008］ and NAFLD remained significant. Elevated levels of phosphatidylcholine （16：0/20：5） ［β=0.026， 95%CI（0.015， 0.036）， P=9.93×10^-7］， phosphatidylcholine （O-16：0/22：5） ［β=0.057， 95%CI（0.035， 0.078）， P=1.79×10^-7］， triacylglycerol （56：7） ［β=0.057， 95%CI （0.035，0.079）， P=2.53×10^-7］ and triacylglycerol （56：8） ［β=0.067， 95%CI （0.047，0.087）， P=1.19×10^-10］ were significantly associated with increased ALT levels. Triacylglycerol （53：2） ［β=0.160， 95% CI （0.123， 0.197）， P=1.55×10^-17］ showed a significant positive causal association with AST levels. Conclusion Specific plasma lipids were causally associated with NAFLD risk and liver enzyme levels， with some associations independent of BMI.

Objective To discuss the expression characteristics of natural autoantibodies against the apoptosis inhibitor gene BIRC5 in plasma of the patients with hepatocellular carcinoma （HCC） and their clinical significances， and to evaluate the potential value as the prognostic biomarker for HCC. Methods A total of 119 HCC patients were enrolled as case group. According to the Barcelona Clinic Liver Cancer （BCLC） staging system， the patients were divided into stages 0+A stage， B stage， and C+D stage. A total of 132 healthy control subjects （healthy control group） were recruited from the physical examination center of our hospital. The blood samples were collected from all the subjects. Two highly efficient linear antigenic peptides （BIRC5a and BIRC5b） for detection were designed and synthesized using epitope prediction software； optimized enzyme-linked immunosorbent assay （ELISA） method was used to detect the levels of BIRC5 natural autoantibodies in plasma of the subjects in two groups； receiver operating characteristic （ROC） curve was plotted to evaluate diagnostic performance； Pearson/Spearman correlation analysis was used to examine the association between antibody levels and clinical indicators ［alpha-fetoprotein （AFP）， liver function， carcinoembryonic antigen （CEA）， carbohydrate antigen 199 （CA199）， total bilirubin （TBil）， albumin （Alb）， and prothrombin time （PT）］. Results Compared with healthy control group， the levels of BIRC5a and BIRC5b natural autoantibodies in plasma of male HCC patients in case group were increased （P<0.05）， while no statistically significant difference was observed in female HCC patients （P>0.05）. Compared with healthy control group， the level of BIRC5a natural autoantibodies in plasma of the patients with BCLC stage C+D in case group was increased （P<0.05）. The correlation analysis results showed that the level of BIRC5a natural autoantibodies increased with higher BCLC stage （r=0.132， P<0.05）， was positively correlated with increased alkaline phosphatase （ALP） and prolonged PT （r=0.328， P<0.05； r=0.247， P<0.05）， and was negatively correlated with decreased Alb （r=-0.423， P<0.01）. The ROC curve analysis results showed that the area under the curve （AUC） for BIRC5a was 0.559； the AUC for BIRC5b was 0.543； the combined analysis AUC was 0.561， and it demonstrated the best diagnostic performance （AUC=0.569） under the optimal diagnostic cut-off value， with a sensitivity of 45.1% and a specificity of 74.6% for diagnosing HCC. Conclusion BIRC5a natural autoantibody is a potential biomarker for HCC prognosis， particularly in male HCC patients. Its elevated plasma level is significantly associated with disease progression and deterioration of liver function， suggesting it may serve as an independent risk factor affecting the prognosis of HCC.

Open bite malocclusion， as a clinically common type of malocclusion， presents complex treatment processes and a high relapse rate， making it one of the challenges in orthodontic treatment. The author of this article reported the diagnosis and treatment process of a teenage patient with a grade Ⅲ open bite to provide reference for the clinicians in managing such conditions. The patient， a 13-year-old male， sought treatment for “misaligned teeth and inability to bite with front teeth” to improve occlusal function and facial appearance. The clinical manifestations included a grade Ⅲ anterior open bite， grade Ⅲ maxillary crowding， and macroglossia. The diagnosis was Angle Class Ⅱ malocclusion and Mao’s ClassⅣ²+Ⅰ¹+Ⅴ malocclusion. A satisfactory treatment outcome was achieved through maxillary single-jaw extraction combined with the use of a lingual arch. After treatment， the patient’s open bite was significantly improved， with well-aligned upper and lower teeth and an intercuspal occlusion relationship. In the treatment of open bite malocclusion， it is essential to correct bad habits as early as possible， leveraging the patient’s growth potential， the wedge effect of posterior teeth， and the pendulum effect of anterior teeth to correct the open bite.

Desmoplastic fibroma （DF） is an extremely rare benign tumor of the jaw bone. Due to its lack of distinct specific features， it is often difficult to distinguish clinically from other diseases， which can lead to suboptimal therapeutic outcomes and increased treatment complexity. This article reportsed the clinical features of an elderly patient with DF to provide the reference for its diagnosis and treatment. The patient， a 66-year-old female， was admitted to the hospital due to recurrent swelling and discomfort in the left posterior mandibular region for over 20 years. The physical examination results showed a hard， tender bony expansion in the left posterior mandible. The panoramic radiography and cone-beam computed tomography （CBCT） results showed an ill-defined radiolucent shadow， about 1.5 cm×1.8 cm， and bone destruction. To confirm the diagnosis， a preoperative pathological biopsy was performed， which showed a spindle cell lesion with a background of abundant fibrous connective tissue. Based on the above findings， the definitive diagnosis of mandibular DF was made. Given the aggressive nature of DF， the lesion was resected via partial mandibulectomy. The defect was repaired using a reconstruction plate for internal fixation and a submandibular gland flap. The postoperative pathology examination results showed dense fibrous connective tissue， with some cellular components exhibiting atypia and the presence of some mast cells， consistent with the pathological characteristics of DF. No postoperative complications or recurrence were observed during the 1-year follow-up period. DF often lacks specific features， which results in the difficulties in diagnosis and treatment. Comprehensive diagnosis requires the integration of imaging and pathological examinations， and treatment involves surgical resection with wide margins. Although DF is typically benign， it exhibits local aggressiveness， necessitating postoperative observation and follow-up for the patients.

46，XX male sex reversal syndrome （SRS） is a rare genetic disorder characterized by abnormal sex development. This article reported the clinical manifestations and auxiliary examination data of a patient with 46，XX male SRS. The patient， an 18-year-old with male social gender， had no history of medication， allergies， or family genetic diseases. He presented to the Reproductive Medicine Center of our hospital due to “mild gynecomastia”. The patient exhibited a male phenotype. The physical examination results revealed absence of beard， an inconspicuous Adam’s apple， absence of axillary hair， mild breast development， sparse pubic hair， and poorly developed external genitalia. The penile length in the flaccid state was approximately 4 cm； bilateral testes were slightly hard in texture， with volumes each approximately 2 mL. The semen analysis results showed that two consecutive routine semen examinations showed no sperm. The sex hormone tests results showed that follicle-stimulating hormone （FSH） 31.41 IU·L^-1， luteinizing hormone （LH） 22.61 IU·L^-1， testosterone （T） 1.04 μg·L^-1， estradiol （E₂） 21.39 ng·L^-1， prolactin （PRL） 16.46 μg·L^-1. The breast color ultrasound results suggested glandular echoes deep to the nipples of both breasts， with the thicker part on the left approximately 1.48 cm and on the right approximately 1.41 cm. The male reproductive system color ultrasound examination results indicated the left testis size was 1.58 cm×1.20 cm×0.77 cm and the right testis size was 1.58 cm×1.26 cm×0.78 cm. Karyotype （high-resolution G-banding 550 bands） was 46，XX. The Y chromosome microdeletion testing results showed deletions in the a+b+c regions of the azoospermia factor （AZF） on the Y chromosome. The detection sites sY84， sY86， sY127， sY134， sY254， and sY255 were all missing. The fluorescence in situ hybridization （FISH） analysis results showed a cryptic translocation of the sex-determining region Y （SRY） gene at the terminus of the short arm of one of the patient’s X chromosomes. The authors of this article discussed the clinical manifestations and auxiliary examination data of this case of 46，XX male SRS， which can provide the reference for improving the clinical understanding， diagnosis， and treatment level of 46，XX male SRS among clinical practitioners.

Spinal gout often involves the facet joints and may form tophi in the spinal canal， but the studies about disc tissue involvement and compressing nerves and causing radicular symptoms in the patients were rare. The clinical materials of one spinal gout patient with intervertebral disc involvement and nerve compression were reported， and the relevant literatures were analyzed. The patient， male， 37-year-old， had low back pain accompanied by right lower limb pain and discomfort for 7 years， aggravated for 2 weeks.The patient had a 10-year history of gout with poorly controlled uric acid levels. The physical examination results revealed mild tenderness over the lower lumbar spinous process and paraspinal area， a positive straight leg raise test on the right side， grade Ⅲ muscle strength in the right iliopsoas muscle， and reduced superficial sensation over the lateral right calf. The uric acid level was 292.2 μmol·L?1. The CT examination results showed bone destruction at the lower endplate of the L5 vertebra and the upper endplate of the S1 vertebra， with scattered high-density shadows within the disc. The magnetic resonance imaging （MRI） results revealed L5 and S1 lumbar disc herniation and irregular signal shadows at the lower endplate of L5 and the upper endplate of S1. The initial diagnosis was “lumbar disc herniation， spinal gout， and gout”. The unilateral biportal endoscopic lumbar discectomy was performed. The pathological examination results of the resected disc tissue confirmed the definite diagnosis of spinal gout. The postoperative symptomatic treatment， including pain relief and uric acid reduction， was administered. At the 3-month postoperative follow-up， the patient’s low back pain and lower limb pain had completely resolved. The manifestations of spinal gout are often similar to those of disc herniation and intraspinal space-occupying lesions， and can include low back pain and lower limb pain. MRI examination in the suspected patients is helpful for the early diagnosis and treatment of spinal gout.

Toll-like receptors （TLRs） are the most well-characterized family of pattern recognition receptors （PRRs）， playing key roles in both innate and adaptive immune processes. In addition to their widespread expression in immune cells， TLRs are also expressed in many tumor cell populations. Their activation can lead to tumor progression or regression， which has made the clinical relevance of TLRs to tumors a research hotspot in recent years. Among them， the TLR subtypes TLR7/8 have dual effects in tumorigenesis and development. On the one hand， they exert tumor-suppressive effects by inducing effective anti-tumor immune responses and mediating tumor cell death； on the other hand， they can also promote tumor progression by regulating tumor cell behavior and the tumor microenvironment （TME）. Therefore， TLR7/8 has become a potential target for tumor prevention and treatment. This article summarizes and analyzes the molecular characteristics and signaling pathways of TLR7/8 and their impact on the process of tumorigenesis and development based on the latest research findings at home and abroad， and systematically reviews the research progress in the application of small molecule modulators targeting TLR7/8， aiming to provide a reference for the discussion on the role of TLR7/8 in tumor prevention and treatment.

Oligodendroglioma （ODG） is a primary glioma that frequently occurs in the frontal lobe of the brain and is associated with a relatively favorable patient prognosis. With technological advancements， the diagnosis of ODG is no longer limited to computed tomography（CT） and magnetic resonance imaging（MRI）， as artificial intelligence and imaging technologies have improved diagnostic accuracy. Pathological and molecular marker diagnostics provide a more precise basis for grading ODG and developing personalized treatment strategies. Currently， significant differences exist in the choice of ODG treatment approaches across different countries or regions. In China， surgical resection remains the core treatment for ODG. Depending on the ODG patients’ specific condition and individual differences after operation， the clinicians can select radiotherapy and/or chemotherapy as adjunctive treatments. Novel therapies such as immunotherapy， tumor-treating fields（TTFields）， and targeted therapy have been applied in clinical practice or are under investigation. Additionally， emerging strategies such as bioactive substance nanocarrier technology and drug repurposing offer new possibilities for treatment. Through the literature review， this article systematically summarized domestic and international diagnostic methods and treatment modalities for ODG， aiming to enhance the precision of its diagnosis and treatment.

Cardiovascular disease （CVD） is one of the leading causes of disability and death worldwide. Its pathological features include irreversible loss of cardiomyocytes （CMs）， cardiac fibrosis， and progressive decline in function. Although existing drug therapies and interventional measures can alleviate the symptoms， they hardly achieve effective regeneration of myocardium tissue， leaving clinical needs far from being met. The bone marrow mesenchymal stem cells （BMSCs）， as an important source of endogenous stem cells， possess characteristics such as proliferation and multidirectional differentiation. In vitro， the BMSCs can be directionally differentiated into the cardiomyocyte-like cells through methods such as chemical reagents， physical stimulation， cytokines， simulation of the myocardial microenvironment， and gene transfection. The differentiation of the BMSCs into the cardiomyocyte-like cells and the repair of damaged CMs primarily rely on their paracrine effects， which also involve the production of exosomes（Exo） via paracrine signaling. The Exo can carry cytokines， phospholipids， and various RNAs， such as microRNAs（miRNA） （miR-29b-3p/miR-125b） and long non-coding RNAs （lncRNAs）. They exert protective effects by regulating targets like a disintegrin and metalloproteinase with thrombospondin motifs 16 （ADAMTS16） and sirtuin 7 （SIRT7）， thereby inhibiting CMs apoptosis and alleviating myocardial fibrosis and inflammatory responses. Furthermore， BMSCs can act on regulatory T lymphocytes （Tregs） in vivo， stimulating them to produce repair factors and promoting macrophage polarization. They also participate in immune responses by modulating natural killer （NK） cells to influence autophagy， thereby mitigating myocardial inflammatory reactions. This article reviews the methods and related mechanisms for the directional differentiation of BMSCs into the cardiomyocyte-like cells in vitro， as well as the protective effects of the BMSCs in CVD models such as diabetic cardiomyopathy （DCM）， myocardial infarction （MI）， and myocardial ischemia-reperfusion injury （MIRI）. It aims to reveal their protective mechanisms on CMs and provide insights for clinical application.

Intertrochanteric femoral fracture （ITFF） refers to a fracture occurring between the greater trochanter and the lesser trochanter， which is an extracapsular fracture of the hip joint. Among these， the type involving medial wall injury is relatively common in clinical practice. The treatment for ITFF involving medial wall injury is primarily surgical. Surgical methods mainly include extramedullary fixation and intramedullary fixation. However， there is currently no unified consensus on which internal fixation method to adopt， especially for complex cases involving medial wall instability， where the choice between extramedullary and intramedullary fixation remains controversial. The optimal treatment approach requires individualized decision-making based on the stability of the medial wall and the patient’s baseline conditions， such as bone quality， complications， and age. Nevertheless， for patients with unstable medial wall injury， priority should be given to restoring the bony support of the medial wall and cortical continuity to reconstruct mechanical stability and reduce the risk of internal fixation failure. This article reviews the anatomical structure of the intertrochanteric region， the injury mechanism of ITFF， fracture classification， biomechanical research， and internal fixation treatment methods for ITFF involving medial wall injury. It systematically compares the advantages and disadvantages of different fixation strategies， aiming to provide the reference for the treatment of ITFF patients with medial wall injury， with the goal of improving patient prognosis and promoting standardized clinical management.