Objective To discuss the inhibitory effect of diosmetin （DIO） on the ferroptosis induced by the glutathione peroxidase （GSH-Px） inhibitor （1S，3R）-RSL3 （RSL3） in spermatocytes GC-2 of the mice， and to clarify the mechanism. Methods The GC-2 cells were divided into control group， RSL3 group， RSL3+0.8 nmol·L?1 DIO group， RSL3+4.0 nmol·L?1 DIO group， RSL3+20.0 nmol·L?1 DIO group， and RSL3+ferroptosis inhibitor Ferrostain-1（Fer-1） group （200 nmol·L?1 Fer-1）. The cells were treated with 0， 1， 5， 10， 50， 100， 500， and 1 000 nmol·L?1 RSL3 solutions， and 0， 0.5， 0.1， 1.0， 5.0， 10.0， and 50.0 μmol·L?1 DIO solutions， respectively. Additionally， the GC-2 cells were divided into blank group， model group， and treatment group. The GC-2 cells in treatment group were further divided into 0.8， 4.0， and 20.0 nmol·L?1 DIO groups， as well as RSL3+0.8 nmol·L?1 DIO group， RSL3+4.0 nmol·L?1 DIO group， and RSL3+20.0 nmol·L?1 DIO group. MTT method was used to detect the survival rates of the GC-2 cells in various groups. The GC-2 cells were treated with 100 nmol·L?1 RSL3 for 0， 6， 12， 24， 36， and 48 h； Western blotting method was used to detect the expression levels of ferroptosis-related proteins in the GC-2 cells in various groups； kits were used to detect the activities of superoxide dismutase （SOD）， levels of malondialdehyde （MDA）， and ratios of glutathione （GSH） to glutathione disulfide （GSSG） in the GC-2 cells in various groups； immunofluorescence method was used to detect the fluorescence intensities of acyl-CoA synthetase long-chain family member 4 （ACSL4） protein in the GC-2 cells in various groups. Results The MTT method results showed that compared with 0 nmol·L^-1 RSL3 group， the survival rates of the GC-2 cells in 50， 100， 500， and 1 000 nmol·L^-1 RSL3 groups were significantly decreased （P<0.01）； compared with 0 μmol·L^-1 DIO group， the survival rates of the GC-2 cells in 0.5， 1.0， 5.0， 10.0， and 50.0 μmol·L^-1 DIO groups were significantly decreased （P<0.01）， and 100 nmol·L^-1 RSL3 with DIO concentration< 0.1 μmol·L^-1 were selected for the subsequent experiments. Compared with blank group， the survival rates of the GC-2 cells in model group was significantly decreased （P<0.01）； compared with model group， the survival rates of the GC-2 cells in RSL3 + 20.0 nmol·L^-1 DIO group was significantly increased （P<0.01）. The Western blotting results showed that compared with 0 h， the expression level of GPX4 protein in the GC-2 cells was significantly decreased after treated with RSL3 for 6 h （P<0.01）， and the expression level of HO-1 protein was significantly increased after treated with RSL3 for 12 h （P<0.05）； after treated with RSL3 for 12 h， the expression levels of GPX4 and FTH1 proteins were significantly decreased （P<0.05 or P<0.01）； after treated with RSL3 for 24 h， the expression levels of GPX4 and HO-1 proteins were significantly decreased （P<0.05 or P<0.01）； after treated with RSL3 for 36 and 48 h， the expression levels of HO-1 protein were significantly decreased （P<0.01）. Therefore， 100 nmol·L^-1 RSL3 and for 12 h were selected as the experimental condition for the subsequent experiments.Compared with control group， the MDA level in the GC-2 cells in RSL3 group was significantly increased （P<0.01）， and the SOD activity and GSH/GSSG ratio were significantly decreased （P<0.05）. Compared with RSL3 group， the SOD activities in the cells in RSL3+0.8 nmol·L^-1 DIO group， RSL3+4.0 nmol·L^-1 DIO group， RSL3+20.0 nmol·L?1 DIO group， and RSL3+Fer-1 group were significantly increased （P<0.05 or P<0.01）. The MDA levels in the cells in RSL3+20.0 nmol·L^-1 DIO group and RSL3+Fer-1 group were significantly decreased （P<0.05 or P<0.01）， and the GSH/GSSG ratio in the cells in RSL3+4.0 nmol·L^-1 DIO group， RSL3+20.0 nmol·L^-1 DIO group， and RSL3+Fer-1 group were significantly increased （P<0.05 or P<0.01）.The immunofluorescence observation results showed that compared with control group， the fluorescence intensity of ACSL4 protein in the GC-2 cells in RSL3 group was significantly increased； compared with RSL3 group， the fluorescence intensities of ACSL4 protein in the cells in RSL3+0.8 nmol·L^-1 DIO group， RSL3+4.0 nmol·L^-1 DIO group， RSL3+20.0 nmol·L^-1 DIO group， and RSL3+Fer-1 group were significantly decreased.The Western blotting results showed that compared with control group， the expression level of HO-1 protein in the cells in RSL3 group was increased （P<0.05）， and the expression levels of GPX4 and FTH1 proteins were significantly decreased （P<0.05 or P<0.01）； compared with RSL3 group， the expression levels of HO-1 protein in the cells in RSL3+0.8 nmol·L^-1 DIO group， RSL3+4.0 nmol·L?1 DIO group， RSL3+20.0 nmol·L^-1 DIO group， and RSL3+Fer-1 group were significantly decreased （P<0.05 or P<0.01）， and the expression levels of GPX4 and FTH1 proteins were significantly increased （P<0.05 or P<0.01）. Conclusion DIO can alleviate the RSL3-induced ferroptosis in the GC-2 spermatocytes of the mice， and its mechanism may be related to the inhibition of HO-1 protein expression and the upregulation of expressions of GPX4 and FTH1 proteins.

Objective To discuss the effect of inhibiting microRNA（miR）-193a-5p expression on pulmonary fibrosis in the rats with acute respiratory distress syndrome（ARDS）， and to clarify the related mechanism. Methods A total of 60 male SD rats were divided into sham operation group， model group， miR-193a-5p antagonist group（Antagomir group）， and negative control group （Antagomir-NC group）， and there were 15 rats in each group. The ARDS animal model was induced by administering 10 mg·kg^-1 lipopolysaccharide（LPS） via tracheal instillation， while the rats in sham operation group received an equal volume of saline. After successful modeling， the rats in Antagomir group and Antagomir-NC group were treated with miR-193a-5p Antagomir or Antagomir-NC via tail vein injection. The arterial partial pressure of oxygen （PaO₂） and oxygenation index（OI） of the rats in various groups were measured； HE staining and Masson staining were used to observe the pathology and collagen fiber deposition in lung tissue of the rats； kit was used to detect the level of hydroxyproline（Hyp） in lung tissue of the rats in various groups； enzyme-linked immunosorbent assay（ELISA） method was used to detect the levels of inflammatory factors tumor necrosis factor α（TNF-α）， interleukin （IL）-1β， and IL-6 in bronchoalveolar lavage fluid（BALF） of the rats in various groups； real-time fluorescence quantitative PCR（RT-qPCR） method was used to detect the expression levels of miR-193a-5p in lung tissue of the rats in various groups； Western blotting method was used to detect the expression levels of β-catenin， Snail family transcriptional repressor 1（Snail1）， and α-smooth muscle actin（α-SMA） proteins in lung tissue of the rats in various groups. Results Compared with sham operation group， the PaO₂ and OI of the rats in model group were significantly decreased （P<0.05）； compared with model group， the PaO₂ and OI of the rats in Antagomir group were significantly increased（P<0.05）. The HE staining results showed that the lung tissue structure of the rats in sham operation group was normal， and there were no obvious inflammatory changes； compared with sham operation group， mild abnormalities in lung tissue structure， alveolar atrophy， and collapse were observed in the rats in model group and Antagomir-NC group， with a large number of lymphocytes and a small number of neutrophils infiltrating in the alveolar cavities， and widened alveolar spaces； compared with model group， the rats in Antagomir group showed a significant reduction in lymphocytes and neutrophil infiltration in the alveolar cavities and there were no obvious hyperplasia. The Masson staining results showed no obvious blue collagen fiber deposition in lung tissue of the rats in sham operation group； compared with sham operation group， significant blue collagen fiber deposition was observed in lung tissue of the rats in model group and Antagomir-NC group， with severe damage of the alveolar structure， indicating obvious pulmonary fibrosis； compared with model group， the deposition of blue-stained collagen fibers in lung tissue of the rats in Antagomir group was significantly reduced. Compared with sham operation group， the level of Hyp in lung tissue of the rats in model group was significantly increased（P<0.05）； compared with model group， the level of Hyp of the rats in Antagomir group was significantly decreased（P<0.05）. The ELISA results showed that compared with sham operation group， the levels of TNF-α， IL-1β， and IL-6 in BALF of the rats in model group were significantly increased（P<0.05）； compared with model group， the levels of TNF-α， IL-1β， and IL-6 of the rats in Antagomir group were significantly decreased（P<0.05）. The RT-qPCR results showed that compared with sham operation group， the expression level of miR-193a-5p in lung tissue of the rats in model group was significantly increased（P<0.05）； compared with model group， the expression level of miR-193a-5p of the rats in Antagomir group was significantly decreased（P<0.05）. The Western blotting results showed that compared with sham operation group， the expression levels of β-catenin， Snail1， and α-SMA proteins in lung tissue of the rats in model group were significantly increased（P<0.05）； compared with model group， the expression levels of β-catenin， Snail1， and α-SMA proteins in lung tissue of the rats in Antagomir group were significantly decreased（P<0.05）. Conclusion Inhibition of miR-193a-5p expression can improve the lung function and alleviate the pulmonary fibrosis in the ARDS rats by reducing the inflammatory responses and downregulating the expressions of β-catenin， Snail1， and α-SMA proteins.

Objective To discuss the effect of Boschnikia rossica polysaccharides rapa polysaccharides （BRPS） on lipopolysaccharide （LPS）-induced inflammatory responses in the THP-1 macrophages， and to clarify its mechanism. Methods The THP-1 monocytes were differentiated into the macrophages， and the inflammation model was established using LPS to induce the THP-1 macrophages. CCK-8 method was used to detect the survial rates of the THP-1 macrophages after treated with different concentrations （0， 100， 200， 500， 1 000， and 2 000 μg·L^-1） of LPS and different concentrations （0， 12.5， 25.0， 50.0， 100.0， and 200.0 mg·L^-1） of BRPS to select the concentrations for the subsequent experiments. The THP-1 macrophages were divided into blank group， model group， low dose of BRPS group （25.0 mg·L^-1 BRPS）， medium dose of BRPS group （50.0 mg·L^-1 BRPS）， and high dose of BRPS group （100.0 mg·L^-1 BRPS）. P38 inhibitor SB203580， ERK inhibitor U0126， c-Jun N-terminal kinase（JNK） inhibitor SP600125， and nuclear factor of kappa B（NF-κB） inhibitor BAY11-7082 were used to verify the effects on THP-1 cells. The THP-1 cells were divided into control group， LPS group， inhibitor group， 100.0 mg·L^-1 BRPS group， and inhibitor+100.0 mg·L^-1 BRPS group. ELISA method was used to detect the levels of tumor necrosis factor α （TNF-α）， interleukin （IL）-6， and IL-1β in culture fluid of the THP-1 macrophages in various groups； DCFH-DA fluorescence probe method was used to detect the reactive oxygen species （ROS） levels in the THP-1 macrophages in various groups； Hoechst33342/PI fluorescence staining method was used to detect the membrane damage in the THP-1 macrophages in various groups； JC-1 fluorescence staining was used to observe mitochondrial membrane potential in the THP-1 macrophages in various groups； Western blotting method was used to detect the expression levels of cyclooxygenase-2 （COX-2）， high mobility group protein B1 （HMGB1）， NOD-like receptor thermal protein domain assciated protein 3 （NLRP3）， cysteinyl aspartate specific protease （Caspase）-1， gasdermin D （GSDMD）-N， IL-1β， mitogen-activated protein kinase （MAPK）， and nuclear factor-kappa B （NF-κB） related proteins in the THP-1 macrophages in various groups. Results The CCK-8 method results showed that when the LPS concentration was 100-2 000 μg·L^-1， the survival rates of the THP-1 macrophages were over 90%. Compared with 0 μg·L^-1 LPS group， the IL-6 levels in culture fluid of the THP-1 macrophages in 100， 200， 500， 1 000， and 2 000 μg·L^-1 LPS group were increased （P<0.05）， indicating a significant enhancement of the inflammatory response in the macrophages， so 100 μg·L^-1 LPS was used to construct the inflammation model.After treated with 12.5， 25.0， 50.0， 100.0， and 200.0 mg·L^-1 BRPS， the survival rates of the THP-1 macrophage were 91.2%， 93.8%， 91.4%， 90.6%， and 91.8%， respectively， so 25.0， 50.0， and 100.0 mg·L^-1 BRPS were selected as the drug concentrations for low， medium， and high doses of BRPS groups in the subsequent experiments.The ELISA results showed that compared with blank group， the levels of IL-6， TNF-α， and IL-1β in culture fluid of the THP-1 macrophages in model group were increased （P<0.05）； compared with model group， the levels of IL-6， TNF-α， and IL-1β in low， medium， and high doses of BRPS groups were decreased （P<0.05）. The DCFH-DA fluorescence probe method results showed that compared with blank group， the ROS level in the THP-1 macrophages in model group was increased （P<0.05）； compared with model group，the ROS levels in low， medium， and high doses of BRPS groups were decreased （P<0.05）. The Hoechst33342/PI fluorescence staining results showed that compared with blank group， the degree of membrane damage in the THP-1 macrophages in model group was increased； compared with model group， the degrees of membrane damage in low， medium， and high doses of BRPS groups were decreased. The JC-1 fluorescence staining results showed that compared with blank group， the mitochondrial membrane potential in the THP-1 macrophages in model group was decreased significantly； compared with model group， the mitochondrial membrane potential in low， medium， and high doses of BRPS groups were increased gradually.The Western blotting results showed that compared with blank group， the expression levels of COX-2， HMGB1， NLRP3， Caspase 1， GSDMD-N， and IL-1β proteins and the ratios of p-P38/P38， p-ERK/ERK， p-JNK/JNK， and p-NF-κB/NF-κB in the THP-1 macrophages in model group were increased （P<0.05）； compared with model group， the expression levels of HMGB1， NLRP3， Caspase-1， GSDMD-N， and IL-1β proteins and the ratios of p-P38/P38， p-ERK/ERK， p-JNK/JNK， and p-NF-κB/NF-κB in the THP-1 macrophages in medium and high doses of BRPS groups were decreased （P<0.05）， the expression levels of NLRP3， Caspase-1， and IL-1β proteins in the cells in low dose of BRPS group were decreased （P<0.05）， the expression level of COX-2 protein in the cells in high dose of BRPS group was decreased （P<0.05）. Compared with control group， the ratios of p-P38/P38， p-ERK/ERK， p-JNK/JNK， and p-NF-κB/NF-κB， and the expression level of IL-1β protein in the THP-1 macrophages in LPS group were increased （P<0.05）； compared with LPS group， the ratios of p-P38/P38， p-ERK/ERK， p-JNK/JNK， and p-NF-κB/NF-κB， and the expression level of IL-1β protein in the THP-1 macrophages in inhibitor group， 100 mg·L^-1 BRPS group， and inhibitor+100 mg·L^-1 BRPS group were decreased （P<0.05）； compared with inhibitor group， the ratios of p-P38/P38， p-ERK/ERK， p-JNK/JNK， and p-NF-κB/NF-κB in the THP-1 macrophages in inhibitor+100 mg·L^-1 BRPS group were decreased （P<0.05）. Conclusion BRPS inhibits the inflammatory response of the THP-1 macrophages， which may be related to the MAPK and NF-κB signaling pathways regulated by BRPS.

Objective To discuss the effect of chrysophanol on hydrogen peroxide （H₂O₂）-induced oxidative damage of the EA.hy926 cells， and to clarify its therapeutic role in bronchopulmonary dysplasia （BPD） and related mechanism. Methods The EA.hy926 cells were induced with 25， 50， 100， 200， 400， 800， and 1 600 μmol·L^-1 H₂O₂， and 8， 16， 32， 64， 128， and 256 μmol·L^-1 chrysophanol. CCK-8 method was used to detect the viabilities of the EA.hy926 cells treated with different concentrations of H₂O₂ and chrysophanol. The cells were divided into control group， model group （200 μmol·L^-1 H₂O₂）， low dose of chrysophanol group （8 μmol·L^-1 chrysophanol and 200 μmol·L^-1 H₂O₂）， and high dose of emodin group （256 μmol·L^-1 chrysophanol and 200 μmol·L^-1 H₂O₂）. Western blotting method was used to detect the expression levels of apoptosis-inducing factor （AIF） protein in the cytoplasm and nucleus in various groups； immunofluorescence staining was used to detect the AIF nuclear translocation in the cells in various groups； kits were used to detect the activities of superoxide dismutase （SOD） and the levels of malondialdehyde （MDA）， cysteinyl aspartate specific proteinase（Caspase）-8， and Caspase-9 in the cells in various groups. Results Under different concentrations of H₂O₂， the viabilities of EA.hy926 cells showed an inverted S-shaped curve， with good cell viability， and the half-maximal inhibitory concentration （IC₅₀） was 261.52 μmol·L^-1. The cell model was induced by 200 μmol·L^-1 H₂O₂ for 24 h. As the increaseing of concentration of chrysophanol， there was no significant change of the viability in the EA.hy926 cells （P>0.05）， and interventions were performed using 8 and 256 μmol·L^-1 chrysophanol. The Western blotting results showed that compared with control group， the expression level of AIF protein in the nucleus in model group was significantly increased （P<0.05）， and the expression level of AIF protein in the cytoplasm was significantly decreased （P<0.05）. Compared with model group， the expression levels of AIF protein in the nucleus in both low and high doses of chrysophanol groups were significantly decreased （P<0.05）， and the expression level of AIF protein in the cytoplasm was significantly increased（P<0.05）. The immunofluorescence staining results showed that AIF was less localized in the nucleus in the cells in control group. Compared with control group， the positive value of AIF nuclear translocation in model group was significantly increased （P<0.05）； compared with model group， the positive values of AIF nuclear translocation in both low and high doses of chrysophanol groups were significantly decreased （P<0.05）. Compared with control group， the activity of SOD in the cells in model group was significantly decreased（P<0.05）， and the level of MDA was significantly increased（P<0.01）. Compared with model group， the activities of SOD in the cells in low and high doses of chrysophanol groups were significantly increased（P<0.05）， and the level of MDA was significantly decreased（P<0.05 or P<0.01）. There were no significant differences in the levels of Caspase-8 and Caspase-9 in the cells among various groups（P>0.05）. Conclusion Chrysophanol improves the H₂O₂-induced apoptosis of the EA.hy926 cells by inhibiting the oxidative stress and AIF nuclear translocation， which may be beneficial for the treatment of BPD.

Objective To discuss their effects of PLA （polylactic acid）/PTMC （polytrimethylene carbonate） and PLA/PTMC-calcium phosphate ［Ca?（PO?）?］ composite porous scaffolds prepared by 3D printing technology on bone marrow mesenchymal stem cells （BMSCs） of the rabbits， and to clarify their application values in bone defect repairment. Methods After mixing the materials， PLA/PTMC and PLA/PTMC-Ca₃（PO₄）₂ filaments were prepared by desktop filament extruder. The scaffolds were designed by CATIA V5-6R2019 modeling software and fabricated using CreatBot F430 3D printer. The chemical structure of the PLA/PTMC-Ca₃（PO₄）₂ scaffold was detected by infrared spectroscopy. In vitro degradation experiments were used to detect the degradation weight loss rates and pH values of the two scaffolds. A contact angle measuring instrument was used to detect the hydrophilicities of the two scaffolds. The BMSCs were extracted from three newborn New Zealand white rabbits （2-5-day-old）； CCK-8 method was used to detect the proliferation activities of the cells co-cultured with two scaffolds， and Alizarin red staining was used to observe the osteogenic differentiation of the cells co-cultured with two scaffolds. Results Infrared spectroscopy confirmed the successful preparation of composite scaffolds containing PLA， PTMC， and β-Ca?（PO?）?. During degradation for 6-14 weeks， compared with PLA/PTMC scaffold， the degradation rates of the PLA/PTMC-Ca₃（PO₄）₂ scaffold in lipase solution and phosphate-buffered saline （PBS） were significantly increased （P<0.05 or P<0.01）. During degradation for 8-14 weeks， compared with PLA/PTMC scaffold， the pH value of the PLA/PTMC-Ca?（PO?）? scaffold in lipase solution was significantly increased （P<0.01）. Compared with PLA/PTMC scaffold， the contact angle of the PLA/PTMC-Ca?（PO?）? scaffold was significantly decreased （P<0.01）. On days 5 and 7 of cell co-culture， compared with PLA/PTMC scaffold， the proliferation activity of the cells co-cultured with PLA/PTMC-Ca₃（PO₄）₂ scaffold was significantly increased （P<0.05 or P<0.01）. After 21 d of co-culture， both scaffolds overlapped with BMSCs and locally formed calcified nodules， which were stained orange by Alizarin red. Compared with PLA/PTMC scaffold， the number of mineralized calcium nodules in the cells co-cultured with PLA/PTMC-Ca?（PO?）? scaffold was increased， with greater density and deeper color. Conclusion The PLA/PTMC-Ca₃（PO₄）₂ composite porous scaffolds containing PLA， PTMC， and β-Ca₃（PO₄）₂ are successfully prepared by 3D printing technology. These scaffolds exhibit good degradation properties and show advantages in biocompatibility， hydrophilicity， and osteogenic induction； they are excellent materials for the bone defect repairment.

Objective To discuss the effect of over-expression of solute carrier family 7 member 5 （SLC7A5） gene on the apoptosis of ovarian granulosa cells of the rats， and to clarify its related mechanism. Methods Four 3-week-old SPF grade SD female rats were used to extract the primary ovarian granulosa cells of the rats. These cells were divided into negative control group （NC group） and follicle-stimulating hormone receptor （FSHR） staining group （FSHR group）. Immunofluorescence staining was used to detect the expressions of FSHR protein in the ovarian granulosa cells of the rats to identify the successful isolation of the primary ovarian granulosa cells of the rats. The ovarian granulosa cells were divided into control group （transfected with empty vector plasmid） and OE-SLC7A5 group （transfected with SLC7A5 over-expression plasmid）. Real-time fluoresscence quantitative PCR（RT-qPCR） and Western blotting methods were used to verify the transfection efficiency of the cells； flow cytometry was used to detect the apoptotic rates and cell cycle percentages of the ovarian granulosa cells in two groups； RT-qPCR method was used to detect the expression levels of SLC7A5， cysteinyl aspartate specific proteinase （Caspase）-3， Caspase-8， and tumor necrosis factor-α （TNF-α） mRNA in the ovarian granulosa cells in two groups； Western blotting method was used to detect the expression levels of SLC7A5， Caspase-3， cleaved Caspase-3， Caspase-8， cleaved Caspase-8， and TNF-α proteins in the ovarian granulosa cells in two groups. Results The fluorescence microscope observation results showed that the ovarian granulosa cells appeared spindle-shaped or irregular and specifically expressed FSHR. No FSHR green fluorescence was observed in NC group， while FSHR green fluorescence expression was observed in FSHR group， indicating successful isolation of primary ovarian granulosa cells of the rats. Compared with control group， the expression levels of SLC7A5 mRNA and protein in the ovarian granulosa cells in OE-SLC7A5 group were significantly increased （P<0.05）， indicating successful transfection of SLC7A5 over-expression plasmid into the ovarian granulosa cells. The flow cytometry results showed that compared with control group， the apoptotic rate of the cells in OE-SLC7A5 group was significantly increased （P<0.05）. Compared with control group， the percentage of the ovarian granulosa cells at S phase in OE-SLC7A5 group was significantly decreased （P<0.05）. The RT-qPCR results showed that compared with control group， the expression levels of TNF-α， Caspase-3， and Caspase-8 mRNA in the ovarian granulosa cells in OE-SLC7A5 group were significantly increased （P<0.05）. The Western blotting results showed that compared with control group， the expression levels of TNF-α， Caspase-8， cleaved Caspase-8， Caspase-3， cleaved Caspase-3， and SLC7A5 proteins in the ovarian granulosa cells in OE-SLC7A5 group were significantly increased （P<0.05）. Conclusion The increased expression of SLC7A5 protein promotes the apoptosis of the granulosa cells by upregulating the expressions of TNF-α， Caspase-8， and Caspase-3 apoptotic pathways.

Objective To discuss the effect of concentrated growth factor （CGF） on the performance of bone marrow mesenchymal stem cells （BMSCs） sheets, and to clarify the role of CGF-containing composite cell sheets（CS） in the bone defect repairment. Methods In in vitro experiments, the BMSCs were isolated and cultured from two 3-week-old SD rats； Alizarin Red S and Oil Red O staining were used to identify the osteogenic and adipogenic capabilities of BMSCs； CGF liquid extracts （CGFe） was prepared from three 3-week-old SD rats. The cells were divided into control group, traditional CS （BMSC-CS） group, and CGF-containing composite CS （CGF/BMSC-CS） group. The morphology of the CS in two groups was observed by HE staining. Alizarin Red and alkaline phosphatase （ALP） staining were used to detect the osteogenic differentiation of the CS in various groups； cell scratch assay was used to detect the migration abilities of the cells in various groups； real-time fluorescence quantitative PCR（RT-qPCR） method was used to detect the mRNA expression levels of ALP, collagen are type 1 (COL-1), Runt-related transcription factor 2 （RUNX2）, and osteocalcin (OCN) in the cells in various groups. In in vivo experiments, 15 SD rats were randomly divided into control group, BMSC-CS group, and CGF/BMSC-CS group; micro computed tomography （Micro-CT） was used to detect the bone formation parameters in skull defects of the rats in various groups; HE staining and Masson staining were used to observe the morphology of skull defect tissue of the rats in various groups. Results The third-generation BMSCs were spindle-shaped, closely arranged, and grew in a vortex cluster. The Alizarin red staining results showed obvious calcium nodules, and the Oil red O staining showed red lipid droplets, confirming the cells’ ability to undergo osteogenic and adipogenic differentiation. The CS were white and semi-transparent, with slightly curled edges. The peeled CS were irregularly curled and wrinkled. Compared with BMSC-CS group, the CS in CGF/BMSC-CS group were whiter, less transparent, significantly increased in thickness and extensibility, less prone to breakage, and had a certain degree of stickiness and plasticity. The HE staining results showed that compared with BMSC-CS group, the number of the cells of CS in CGF/BMSC-CS group was increased, with denser arrangement and more abundant extracellular matrix (ECM), which wrapped and connected the cells to form an integral sheet-like structure. The Alizarin red and ALP staining results showed that compared with control group, the ALP activity and mineralization uplift value of CS in BMSC-CS group were significantly increased （P<0.05）； compared with control group and BMSC-CS group, the number of osteoblasts and red mineralized nodules in the CS in CGF/BMSC-CS group was significantly increased, with obvious deepening of the staining, increased positive area, and the ALP activity and mineralization uplift value were significantly increased （P<0.05）. Compared with BMSC-CS group， the ALP activity and mineralization uplif value of the CS in CGF/DMSC-CS group were increased （P<0.05）. The cell scratch assay results showed that after 24 of culture, compared with control group, the migration rates of the cells in BMSC-CS group and CGF/BMSC-CS group were significantly increased (P<0.05). Compared with BMSC-CS group, the migration rate of the cells in CGF/BMSC-CS group was significantly increased (P<0.01). After 48 h of culture, compared with control group, the migration rate of the cells in CGF/BMSC-CS group was significantly increased （P<0.05). The RT-qPCR results showed that compared with control group, the expression levels of COL-1 and OCN mRNA in the cells in BMSC-CS group were significantly increased (P<0.01), and the expression levels of ALP, COL-1, OCN, and RUNX2 mRNA in the cells in CGF/BMSC-CS group were significantly increased (P<0.01). Compared with BMSC-CS group, the expression levels of ALP, COL-1, and OCN mRNA in the cells in CGF/BMSC-CS group were significantly increased(P<0.01). The Micro-CT detection results showed that in control group, the boundary of the rat skull defect area was clear, with almost no new bone formation. In BMSC-CS group, a small amount of new bone formed only at the edge of the bone defect in skull of the rats, with a significant gap in the central area of the defect. In CGF/BMSC-CS group, new bone formed along the edge of the bone defect towards the central area in skull of the rats, repairing most of the bone defect. Compared with control group, the bone volume (BV) and trabecular number (Tb.N) of the rats in BMSC-CS group were significantly increased (P<0.05)； the bone volume (BV), bone volume fraction [BV/tissue volume (TV)], trabecular thickness (Tb.Th), and trabecular number (Tb.N) in skull of the rats in CGF/BMSC-CS group,were significantly increased (P<0.05). Compared with BMSC-CS group, the BV, BV/TV, Tb.Th, and Tb.N in skull of the rats in CGF/BMSC-CS group were significantly increased (P<0.01). The HE and Masson staining observation showed that in control group, almost no new bone formed in the skull defect tissue of the rats, with only a large amount of collagen fibers connecting the two sides of the bone ends. In BMSC-CS group, a small amount of new bone formed only at the edge of the bone defect in skull tissue of the rats, with the central area of the defect containing dense collagen fibers connected to the newly formed bone at the defect edge. In CGF/BMSC-CS group, new bone tissue could be seen at the edge of the bone defect, and bone islands formed in the central area of the defect, surrounded by osteocytes and a large amount of collagen fibers. The Masson staining observation results showed that the cytoplasm and osteoid were red, and the collagen was blue. In CGF/BMSC-CS group, newly formed osteoid was observed in skull defect tissue of the rats, with the highest amount of new bone formation. Conclusion CGF can promote the osteogenic differentiation and increase the richness of ECM in BMSCs sheets. CGF-containing composite CS can efficiently repair skull defects of the rats and serve as an ideal and safe material for promoting the bone regeneration.

Objective To discuss the therapeutic effect of resveratrol on the temporomandibular joint osteoarthritis （TMJOA）， and to clarify the related mechanism. Methods Forty-five SD rats were randomly divided into control group， model group， and resveratrol group， and there were 15 rats in each group. The rats in model group and resveratrol group were intra-articularly injected with 50 μL of 20 g·L^-1 monosodium iodoacetate （MIA） to set TMJOA rat models， while the rats in control group were injected with an equal volume of normal saline. Three weeks after modeling， the rats in resveratrol group received an injection of 80 μL resveratrol solution， once a week for three weeks， while the rats in control and model groups were injected with an equal volume of normal saline. Micro-computed tomography （Micro-CT） system was used to detect the condyle structure and the bone volume fraction （BV/TV）， trabecular thickness （Tb.Th）， trabecular spacing （Tb.Sp）， and trabecular number （Tb.N） of the rats in various groups were calculated； HE staining and toluidine blue staining were used to observe the pathomorphology of temporomandibular joint （TMJ） tissue of the rats in various groups； immunohistochemistry was used to detect the expression levels of SRY-related HMG box （SOX）-9， matrix metalloproteinase （MMP）-13， silent information regulator （Sirt）1， phosphatidylinositol 3-kinase （PI3K）， phosphorylated protein kinase B （p-Akt）， and phosphorylated mammalian target of rapamycin （p-mTOR） in TMJ tissue of the rats in various groups； real-time quantitative PCR （RT-qPCR） method was used to detect the expression levels of SOX-9， MMP-13， Sirt1， PI3K， mTOR， and Akt mRNA in TMJ tissue of the rats in various groups. Results Three weeks after modeling， condylar bone was destructed， the surface was roughness， and continuity interruption were observed， indicating TMJOA model of the rats was established successfully. The Micro-CT system results showed that the condylar surface of the rats in control group was smooth and regularly shaped， with continuous bone texture； the rats in model group had significant condylar destruction， disrupted continuity， surface roughness， and varying degrees of bone defects； the rats in resveratrol group showed alleviated condylar lesions and improved appearance. Compared with control group， the BV/TV and Tb.Th of the rats in model group were significantly decreased （P<0.05）， and Tb.Sp was significantly increased （P<0.05）； compared with model group， the BV/TV and Tb.Th of the rats in resveratrol group were significantly increased （P<0.05）， and the Tb.Sp was significantly decreased （P<0.05）. The HE staining results showed clear layers and orderly chondrocyte arrangement in condyle of the rats in control group； the rats in model group showed rough uneven surface， obvious defects， and typical TMJOA features； the rats in resveratrol group showed slightly rough surface with generally clear layers and orderly arranged cells. The toluidine blue staining results showed distinct blue-purple staining of chondrocytes in hypertrophic layer of the rats in control group； pale staining or even loss of staining in some areas of the rats in model group； and distinct and relatively uniform staining in hypertrophic layer of the rats in resveratrol group. The immunohistochemistry results showed that compared with control group， the expression levels of MMP-13， PI3K， p-Akt， and p-mTOR proteins in TMJ tissue of the rats in model group were significantly increased （P<0.05）， while the expression levels of SOX-9 and Sirt1 proteins in TMJ tissue of the rats were significantly decreased （P<0.05）； compared with model group， the expression levels of SOX-9 and Sirt1 proteins in TMJ tissue of the rats in resveratrol group were significantly increased （P<0.05）， whereas the expression levels of MMP-13， PI3K， p-Akt， and p-mTOR proteins were significantly decreased （P<0.05）.The RT-qPCR results showed that compared with control group， the expression levels of MMP-13， PI3K， Akt， and mTOR mRNA in TMJ tissue of the rats in model group were significantly increased （P<0.05）， while the expression levels of SOX-9 and Sirt1 mRNA were significantly decreased （P<0.05）； compared with model group， the expression levels of SOX-9 and Sirt1 mRNA in TMJ tissue of the rats in resveratrol group were significantly increased （P<0.05）， whereas the expression levels of MMP-13， PI3K， Akt， and mTOR mRNA were significantly decreased （P<0.05）. Conclusion Resveratrol has therapeutic effect on TMJOA， and its mechanism may be related to the activation of Sirt1 and inhibition of the PI3K-Akt-mTOR signaling pathway.

Objective To investigate the effects of bisphenol A （BPA） on the proliferation activity and stemness characteristics of endometrial mesenchymal stem/stromal cells （eMSCs）， and to elucidate the improvement effect of human umbilical cord mesenchymal stem cell-derived supernatant （hUCMSC-Sup） on the cell injury. Methods The eMSCs were cultured in vitro and treated with different concentrations of BPA （0， 200， 250， 300， 350， and 400 μmol·L^-1）. The eMSCs were divided into control group（only cultured with culture solution）， BPA group （cultured with isovolumetric culture solution including 200 μmol·L^-1 BPA）， BPA+hUCMSC-Sup group （cultured with isovolumetric culture solution including 200 μmol·L^-1 BPA and 50% volumetric ratio of hUCMSC-Sup）， and BPA+CHIR-99021 group （cultured with isovolumetric culture solution including 200 μmol·L^-1 BPA and 10 μmol·L^-1 CHIR-99021）.The survival rates of eMSCs in various groups were detected by methyl thiazolyl tetrazolium（MTT） assay. The numbers and diameters of the spheroids in various groups were detected by spheroids formation assay， the proliferation activities of the cells in eMSCs stem cell spheroids in various groups were detected by CCK-8 assay； the percentage of CD73+ cells in eMSCs in various groups were detected by flow cytometry； the expression levels of sex determining region Y-box 2 （Sox2）， octamer-binding transcription factor 4（Oct4）， and Nanog mRNA in the eMSCs in various groups were detected by real-time fluorescence quantitative PCR（RT-qPCR） method， the expression levels of β-catenin protein in the eMSCs in various groups were detected by Western blotting method. Results The MTT results showed that after treated with BPA for 24 and 48 h， compared with 0 μmol·L^-1 BPA group， the survival rates of eMSCs in 200， 250， 300， 350，and 400 μmol·L^-1 BPA groups were significantly decreased （P<0.01）. At 24 and 48 h after treatment， compared with control group， the survival rate of the eMSCs in BPA group was significantly decreased （P<0.01）； at 48 h after treatment， compared with BPA group， the survival rate of the eMSCs in BPA+hUCMSC-Sup group was significantly inereased （P<0.05）. The spheroids formation assay results showed that compared with culture 3 d group， the numbers and diameters of stem cell spheroids of the eMSCs in culture 4 d group and culture 5 d group were significantly increased （P<0.05 or P<0.01）； compared with control group， after 48 h of culture， the number and diameter of the cells in eMSCs stem cell spheroids in BPA group were significantly decreased （P<0.05 or P<0.01）. The CCK-8 results showed that after 24 and 48 h of treatment， compared with control group， the proliferation activity of the cells in eMSCs stem cell spheroids in BPA group was significantly decreased （P<0.01）； compared with BPA group， the proliferation activity of the cells in eMSCs stem cell spheroids in BPA+hUCMSC-Sup group was significantly increased（P<0.01）. The flow cytometry results showed that compared with control group， the percentage of the CD73+ cells in the eMSCs in BPA group was significantly decreased （P<0.01）； compared with BPA group， the percentage of the CD73+ cells in eMSCs in BPA+hUCMSC-Sup group was significantly increased （P<0.01）. The RT-qPCR results showed that compared with control group， the expression levels of Sox2， Oct4， and Nanog mRNA in the cells in BPA group were significantly decreased （P<0.01）； compared with BPA group， the expression levels of Sox2， Oct4， and Nanog mRNA in the cells in BPA+hUCMSC-Sup group and BPA+CHIR-99021 group were significantly increased （P<0.01）. The Western blotting results showed that compared with control group， the expression level of β-catenin protein in the eMSCs in BPA group was significantly decreased（P<0.01）； compared with BPA group， the expression levels of β-catenin protein in the eMSCs in BPA+hUCMSC-Sup group and BPA+CHIR-99021 group were signifrcantly inereased （P<0.01）. Conclusion BPA can inhibit the stemness characteristics of the eMSCs， and injury the self-renewal and repair of endometrium； its mechanism may be related to down-regulating the activity of Wnt/β-catenin signal pathway in the cells. hUCMSC-Sup can promote the proliferation of injured eMSCs， and has improvement effect on the stemness injury induced by BPA.

Objective To discuss the protective effect of ginsenoside Rh1 （G-Rh1） on kidney injury in the diabetic mellitus（DM） mice， and to clarify its mechanism. Methods The diabetic kidney disease （DKD） model was prepared by using the high-fat， high-sugar diet combined with intraperitoneal injection of streptozotocin （STZ）. A total of 48 C57/BL6 model mice were randomly divided into model group， nuclear factor erythroid 2-related factor 2 （Nrf2） inhibitor ML385 group （ML385 group） （30 mg·kg^-1）， G-Rh1 group （30 mg·kg^-1）， and G-Rh1+ML385 group （30 mg·kg^-1 G-Rh1+30 mg·kg^-1 ML385）， and there were 12 mice in each group. Additionally， 12 C57/BL6 mice were selected as control group. After treated for 8 weeks， automatic analyzer was used to detect the levels of fasting blood glucose （FBG）， blood urea nitrogen （BUN）， and serum creatinine （Scr） in serum of the mice in various groups， as well as 24 h urinary protein （24 h UP） levels in urine， and the kidney index was calculated； kits were used to detect the activities of superoxide dismutase （SOD） and lactate dehydrogenase （LDH）， and the levels of malondialdehyde （MDA） in kidney tissue of the mice in various groups； Western blotting method was used to detect the expression levels of Nrf2 and heme oxygenase-1 （HO-1） proteins in kidney tissue of the mice in various groups. Results Compared with control group， the levels of FBG and kidney indexes in serum of the mice in model group， ML385 group， and G-Rh1+ML385 group were significantly increased （P<0.01）， and the level of FBG in serum of the mice in G-Rh1 group was significantly increased（P<0.01）； compared with model group， the kidney index of the mice in ML385 group was significantly increased （P<0.05）， while the levels of FBG and kidney index of the mice in G-Rh1 group were significantly decreased （P<0.05 or P<0.01）； compared with G-Rh1 group， the level of FBG and kidney index of the mice in G-Rh1+ML385 group were significantly increased （P<0.01）. Compared with control group， the levels of BUN and Scr in serum， and 24 h UP in urine of the mice in model group， ML385 group， G-Rh1 group， and G-Rh1+ML385 group were significantly increased （P<0.01）； compared with model group， the level of BUN in serum and 24 h UP in urine of the mice in ML385 group were significantly increased （P<0.05）， while the levels of BUN and Scr in serum， and 24 h UP in urine of the mice in G-Rh1 group were significantly decreased （P<0.01）； compared with G-Rh1 group， the levels of BUN and Scr in serum， and 24 h UP in urine of the mice in G-Rh1+ML385 group were significantly increased （P<0.01）. Compared with control group， the activities of SOD in kidney tissue of the mice in model group， ML385 group， G-Rh1 group， and G-Rh1+ML385 group were significantly decreased （P<0.01）， while the levels of MDA and LDH activities were significantly increased （P<0.01）； compared with model group， the activity of SOD in kidney tissue of the mice in ML385 group was significantly decreased （P<0.05）， and the level of MDA was significantly increased （P<0.05）； the activity of SOD in kidney tissue of the mice in of G-Rh1 group was significantly increased （P<0.01）， and the level of MDA and activity of LDH were significantly decreased （P<0.01）； compared with G-Rh1 group， the activity of SOD in kidney tissue of the mice in G-Rh1+ML385 group was significantly decreased （P<0.01）， and the level of MDA and activity of LDH were significantly increased （P<0.01）. Compared with control group， the expression levels of Nrf2 and HO-1 proteins in kidney tissue of the mice in model group， ML385 group， G-Rh1 group， and G-Rh1+ML385 group were significantly decreased （P<0.05 or P<0.01）； compared with model group， the expression levels of Nrf2 and HO-1 proteins in kidney tissue of the mice in ML385 group and G-Rh1+ML385 group were significantly decreased （P<0.05）， while the expression levels of Nrf2 and HO-1 proteins in kidney tissue of the mice in G-Rh1 group were significantly increased （P<0.01）； compared with G-Rh1 group， the expression levels of Nrf2 and HO-1 proteins in kidney tissue of the mice in G-Rh1+ML385 group were significantly decreased （P<0.01）. Conclusion Ginsenoside Rh1 reduces the oxidative stress and improves the kidney function， providing protective effects on kidney injury in the DM mice， and its mechanism may be related to the activation of the Nrf2/HO-1 signaling pathway.

Objective To discuss the effect of curcumin on the proliferation， migration， and invasion of the human prostate cancer C4-2 and LNCaP cells， and to clarify its possible mechanism. Methods The lentiviral transfection system was used to transfect the C4-2 and LNCaP cells， regarded as shCD147-C4-2 group and shCD147-LNCaP group. RNA interference technology was used to prepare the CD147-silenced cells； the cells transfected with an empty vector were regarded as negative control and divided into shNC-C4-2 group （shNC-C4-2 cells） and shNC-LNCaP group （shNC-LNCaP cells）. The C4-2 and LNCaP cells at logarithmic growth phase， as well as shCD147-C4-2 and shCD147-LNCaP cells， were treated with 20 μmol·L^-1 curcumin.The morphology of the cells in various groups was observed under microscope at 0 and 24 h of treatment； MTT method was used to detect the proliferation activities of the cells in various groups； cell scratch assay was used to detect the migration rates of the cells in various groups；Western blotting method was used to detect the expression levels of apoptosis， invasion， and migration-related proteins in the cells in various groups. Results Compared with C4-2 group， the expression of CD147 protein in the cells in shCD147-C4-2 group was significantly decreased after CD147 gene silenting.Compared with LNCaP group， the expression level of CD147 protein in the cells in shCD147-LNCaP group was significantly decreased after CD147 gene silenting. Compared with 0 h of treatment， some cells in C4-2 and LNCaP groups after 24 h of treatment with 20 μmol·L^-1 curcumin， showed apoptosis signs with the presence of typical apoptotic bodies. The apoptotic phenomena in shCD147-C4-2 and shCD147-LNCaP groups was reduced.The MTT assay results showed that compared with C4-2+0 μmol·L^-1 curcumin group， the proliferation activities of the cells in C4-2+20 μmol·L^-1 curcumin group， C4-2+40 μmol·L^-1 curcumin group， C4-2+60 μmol·L^-1 curcumin group， and C4-2+80 μmol·L^-1 curcumin group were decreased （P<0.01）. Compared with LNCaP+0 μmol·L^-1 curcumin group， the proliferation activity of the cells in LNCaP+20 μmol·L^-1 curcumin group， LNCaP+ 40 μmol·L^-1 curcumin group， LNCaP+60 μmol·L^-1 curcumin group， and LNCaP+80 μmol·L^-1 curcumin group were decreased （P<0.01）. Compared with shNC-C4-2 group， the proliferation activity of the cells in shNC-C4-2+20 μmol·L^-1 curcumin group was decreased （P<0.01）. Compared with shNC-C4-2+20 μmol·L^-1 curcumin group， the proliferation activity of the cells in shCD147-C4-2+20 μmol·L^-1 curcumin group was increased （P<0.01）. Compared with shNC-LNCaP group， the proliferation activity of the cells in shNC-LNCaP+20 μmol·L^-1 curcumin group was decreased （P<0.01）； compared with shNC-LNCaP+20 μmol·L^-1 curcumin group， the proliferation activity of the cells in shCD147-LNCaP+20 μmol·L^-1 curcumin group was significantly increased （P<0.01）. The cell scratch healing assay results showed that compared with C4-2 group， the migration rates of the cells in C4-2+20 μmol·L^-1 curcumin group and C4-2+40 μmol·L^-1 curcumin group after 24 h of treatment were decreased （P<0.01）； compared with LNCaP group， the migration rates of the cells in LNCaP+20 μmol·L^-1 curcumin group and LNCaP+40 μmol·L^-1 curcumin group were increased（P<0.01）； compared with shNC-C4-2 group， the migration rate of the cells in shNC-C4-2+20 μmol·L^-1 curcumin group was decreased （P<0.01）； compared with shNC-C4-2+20 μmol·L^-1 curcumin group， the migration rate of the cells in shCD147-C4-2+20 μmol·L^-1 curcumin group was significantly increased （P<0.05）； compared with shNC-LNCaP group， the migration rate of the cells in shNC-LNCaP+20 μmol·L^-1 curcumin group was decreased （P<0.01）； compared with shNC-LNCaP+20 μmol·L^-1 curcumin group， the garation rate of the cells in shCD147-LNCaP+20 μmol·L^-1 curcumin group was significantly increased （P<0.05）.The Western blotting results showed that compared with C4-2 group， the expression levels of Bcl-2-associated X protein （Bax）， cleaved Caspase-3， and poly ADP-ribose polymerase 1 （PARP1） proteins in the cells in C4-2+20 μmol·L^-1 curcumin group and C4-2+40 μmol·L^-1 curcumin group were significantly increased （P<0.01）， and the expression levels of Bcl-2 protein was significantly decreased （P<0.05 or P<0.01）； compared with LNCaP group， the expression levels of Bax， cleaved Caspase-3， and PARP1 proteins in the cells in LNCaP+20 μmol·L^-1 curcumin group and LNCaP+40 μmol·L^-1 curcumin group were significantly increased （P<0.01）， and the expression level of Bcl-2 protein in the cells in LNCaP+40 μmol·L^-1 curcumin group was decreased （P<0.01）； compared with shNC-C4-2 group， the expression levels of Bax， cleaved Caspase-3， and PARP1 proteins in the cells in shNC-C4-2+20 μmol·L^-1 curcumin group were significantly increased （P<0.05 or P<0.01）， and the expression level of Bcl-2 protein was significantly decreased （P<0.05）； compared with shNC-C4-2+20 μmol·L^-1 curcumin group， the expression levels of Bax and cleaved Caspase-3 proteins in the cells in shCD147-C4-2+20 μmol·L^-1 curcumin group were significantly decreased （P<0.01）； compared with shNC-LNCaP group， the expression levels of Bax， cleaved Caspase-3， and PARP1 proteins in the cells in shNC-LNCaP+20 μmol·L^-1 curcumin group were significantly increased （P<0.05 or P<0.01）， and the expression level of Bcl-2 protein was significantly decreased （P<0.05）； compared with shNC-LNCaP+20 μmol·L^-1 curcumin group， the expression levels of Bax， cleaved Caspase-3， and PARP1 proteins in the cells in shCD147-LNCaP+20 μmol·L^-1 curcumin group were significantly decreased （P<0.05 or P<0.01）， and the expression level of Bcl-2 protein was significantly increased （P<0.05）. Compared with C4-2 group， the expression levels of E-cadherin protein in the cells in C4-2+20 μmol·L^-1 curcumin group and C4-2+40 μmol·L^-1 curcumin group were significantly increased （P<0.01）， and the expression levels of N-cadherin and Vimentin proteins were significantly decreased （P<0.01）； compared with LNCaP group， the expression levels of E-cadherin protein in the cells in LNCaP+20 μmol·L^-1 curcumin group and LNCaP+40 μmol·L^-1 curcumin group were significantly increased （P<0.01）， and the expression levels of N-cadherin and Vimentin proteins in the cells in LNCaP+40 μmol·L^-1 curcumin group were significantly decreased （P<0.01）； compared with shNC-C4-2 group， the expression levels of N-cadherin and Vimentin proteins in the cells in shNC-C4-2+20 μmol·L^-1 curcumin group were significantly decreased （P<0.01）； compared with shNC-C4-2+20 μmol·L^-1 curcumin group， the expression level of E-cadherin protein in the cells in shCD147-C4-2+20 μmol·L^-1 curcumin group was significantly decreased（P<0.01）， and the expression levels of N-cadherin and Vimentin proteins were significantly increased （P<0.01）； compared with shNC-LNCaP group， the expression level of E-cadherin protein in the cells in shNC-LNCaP+20 μmol·L^-1 curcumin group was significantly increased （P<0.01）， and the expression levels of N-cadherin and Vimentin proteins were significantly decreased （P<0.01）； compared with shNC-LNCaP+20 μmol·L-1 curcumin group， the expression level of E-cadherin protein in the cells in shCD147-LNCaP+20 μmol·L^-1 curcumin group was significantly decreased （P<0.01）， and the expression level of N-cadherin was significantly increased （P<0.05）. Conclusion Curcumin inhibits the proliferation， migration， and invasion of the prostate cancer cells in vitro and induces the apoptosis； silencing the CD147 gene partially reduces its inhibitory effect and its ability to induce the apoptosis.

Objective To discuss the promotion effect of chemokine C-C motif ligand 19 （CCL19） induced macrophage M1 polarization on chronic pancreatitis of the mice， and to clarify its related mechanism. Methods Ten male C57BL/6N mice were selected， and the pancreatic acinar cells and peritoneal macrophages were extracted from these mice to construct the macrophage-acinar cell co-culture system. The co-culture system cells were divided into control group， model group， and small interfering RNA CCL19 （si-CCL19） group. The morphology of the acinar cells in various groups were observed under microscope. Forty mice were randomly selected and divided into normal group and chronic pancreatitis group， and there were 20 mice in each group. HE staining was used to observe the pathomorphology of pancreatic tissue of the mice in two groups； immunofluorescence staining was used to observe the expressions of cytokeratin 19 （CK19）， amylase， M1 macrophage-related markers inducible nitric oxide synthase （iNOS）， and F4/80 in pancreatic tissue of the mice in two groups and morphology of follicular cells and the expressions of CK19， amylase in the co-culture system cells in various groups； enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of tumor necrosis factor-α （TNF-α）， interleukin （IL）-6， and IL-1β in serum of the mice in two groups and in the co-culture system cells in various groups； immunohistochemistry was used to observe the expression of CCL19 protein in pancreatic tissue of the mice in two groups； Western blotting method was used to detect the expression levels of CCL19 protein and two nuclear factor-κB （NF-κB） signaling pathway-related proteins P65， phosphorylate P65 （p-P65）， kappa B inhibitor of kinase α/β（IKKα/β）， phosphorylated IKKα/β （p-IKKα/β）， IkBα， phosphorylated IκBα（p-IκBα） in pancreatic tissue of the mice in two groups and in the co-culture system cells in various groups. Results The HE staining results showed that the acinar cells in pancreatic tissue of the mice in normal group were tightly arranged； compared with normal group， the acinar cells of the mice in chronic pancreatitis group showed obvious vacuolation and acinar cell ductal metaplasia， indicating successful preparation of the mouse pancreatitis model. The immunofluorescence staining results showed that compared with control group， the acinar cells in model group exhibited severe vacuolation， the CK19 expression was significantly increased， and the amylase expression was significantly decreased； compared with model group， the acinar cell ductal metaplasia in si-CCL19 group was decreased， the CK19 expression was significantly decreased， and the amylase expression was significantly increased； compared with normal group， the expression of amylase in pancreatic tissue of the mice in chronic pancreatitis group was significantly decreased， while the expressions of CK19 and M1 macrophage markers iNOS and F4/80 were significantly increased. The ELISA results showed that compared with normal group， the serum levels of TNF-α， IL-6， and IL-1β of the mice in chronic pancreatitis group were significantly increased （P<0.05）； compared with control group， the levels of TNF-α， IL-6， and IL-1β in the cells in model group were significantly increased （P<0.05）； compared with model group， the levels of TNF-α， IL-6， and IL-1β in the cells in si-CCL19 group were significantly decreased （P<0.05）. The immunohistochemistry results showed that compared with normal group， the expression of CCL19 protein in pancreatic tissue of the mice in chronic pancreatitis group was significantly increased. The Western blotting results showed that compared with normal group， the expression levels of CCL19 protein and NF-κB signaling pathway-related proteins p-P65， p-IKKα/β， and p-IκBα in pancreatic tissue of the mice in chronic pancreatitis group were significantly increased（P<0.05）； compared with control group， the expression levels of CCL19， p-IKKα/β， p-P65， and p-IκBα proteins in the cells in model group were significantly increased （P<0.05）； compared with model group， the expression levels of CCL19， p-IKKα/β， p-P65， and p-IκBα proteins in the cells in si-CCL19 group were decreased （P<0.05）. Conclusion CCL19 promotes the macrophage M1 polarization through the NF-κB signaling pathway， induces the formation of inflammatory microenvironment， and promotes the occurrence and development of pancreatitis.

Objectives To discuss the inhibitory effect of Lactobacillus reuteri on the replication of rotavirus （RV） strain SA11 in vivo and in vitro， and to clarify its effect on the expression of related immune factors. Methods For in vitro experiments， Lactobacillus reuteri was cultured and identified， and the standard curve and growth curve were plotted to screen the optimal time and concentration for Lactobacillus reuteri cultivation. The cells were infected with Lactobacillus reuteri at the concentrations of 5×10⁸， 10×10⁸， 50×10⁸， 100×10⁸， 200×10⁸， and 500×10⁸ CFU·mL^-1， and the surival rates of Caco-2 cells were detected by trypan blue staining method. Various concentrations of Lactobacillus reuteri were co-incubated with RV in vitro and applied to the Caco-2 cells. The cells were divided into negative control group （NC group）， positive control group （PC group）， and 10⁷， 10⁸， 10⁹， and 10¹⁰ CFU·mL^-1Lactobacillus reuteri groups. Immunofluorescence focus method was used to detect the viral titers in the Caco-2 cells after treated with Lactobacillus reuteri and real-time fluorescence quantitative PCR（RT-qPCR） method was used to detect the copy numbers of RV VP6 gene in the Caco-2 cells after treated with various concentrations of Lactobacillus reuteri. In in vivo experiments， 25 litters of SPF suckling mice were divided into control group， RV group （infected with SA11 strain）， Ab-NC group （treated with antibiotic to deplete gut microbiota）， Ab-RV group （depleting gut microbiota and then infected with SA11 strain）， and Ab-Lac-RV group （depleting gut microbiota， treated with Lactobacillus reuteri， and then infected with SA11 strain）. The fecal samples were collected on days 2， 4， 6， 8， and 10 gavage， colon tissue sample were collected on day 4 of and RT-qPCR method was used to detect the copy numbers of RV VP6 gene in feces and the mRNA expression levels of interleukin （IL）-1β， IL-8， IL-10， interferon-γ （IFN-γ）， and tumor necrosis factor-α （TNF-α） in colon tissue of the suckling mice in vartious groups. Results The Lactobacillus reuteri grew well， with round， smooth， and milky white convex colonies and neat edges. After Gram staining， the bacteria appeared purple， irregular， and square-shaped rods. 16SrDNA sequencing showed 99% sequence homology， indicating successful activation of Lactobacillus reuteri. The number of live Lactobacillus reuteri was linearly related to the absorbance （A） value， and the standard curve for regression analysis was Y=0.437 5X+0.000 6， R²=0.999 4. During the 0-2 h cultivation period， the bacteria were at the logarithmic growth phase with slow growth； from 2-14 h， the bacteria grew rapidly and stabilized at 14-16 h， reaching the growth rate peak at 16 h， after which they entered the decline phase. Infection with Lactobacillus reuteri at concentrations of 5×10⁸， 10×10⁸， 50×10⁸， 100×10⁸， and 200×10⁸ CFU·mL^-1 resulted in the survival rates of Caco-2 cells were all >90%， so these concentrations were selected for the further experiments. Compared with PC group， the copy numbers of RV VP6 gene in the Caco-2 cells in 5×10⁸， 10×10⁸， 50×10⁸， 100×10⁸， and 200×10⁸ CFU·mL^-1Lactobacillus reuteri groups were significantly decreased （P<0.01）. Compared with PC group， the viral titers in the Caco-2 cells in 10⁷， 10⁸， 10⁹， and 10¹⁰ CFU·mL^-1Lactobacillus reuteri groups were significantly decreased （P<0.01）. Compared with control group， the numbers of gut microbiota colonies in Ab-NC， Ab-RV， and Ab-Lac-RV groups were significantly decreased， indicating successful depletion of gut microbiota in the suckling mice. On days 2 and 4 after gavage， the RV VP6 gene copy number in the feces in Ab-RV group was significantly lower than that in RV group （P<0.05）. On days 4， 6， 8， and 10 after gavage， the RV VP6 gene copy number in the feces in Ab-Lac-RV group was significantly lower than that in Ab-RV group （P<0.05）. Compared with control group， the expression levels of IL-1β， IL-10， IFN-γ， and TNF-α mRNA in colon tissue in Ab-RV and Ab-Lac-RV groups were significantly increased （P<0.05 or P<0.01）， while the expression level of IL-8 mRNA was significantly decreased （P<0.05），and the expression level of IL-10 mRNA in colon tissue in Ab-LAC-RV group was significantly increased （P<0.01）. Conclusion Lactobacillus reuteri may inhibit the RV replication by upregulating the expressions of IL-1β， IL-10， IFN-γ， and TNF-α mRNA and downregulating the expression of IL-8 mRNA.

Objective To discuss the effect of nucleoredoxin （NXN） in medial prefrontal cortex （mPFC） region of the mice with post-stroke depression （PSD）， and to clarify its possible mechanism. Methods A total of 42 mice among 80 C57BL/6 mice were randomly divided into NXN over-expression adeno-associated virus infection group （AAV-NXN-OE group， n=21） and negative control adeno-associated virus infection group （AAV-NC group， n=21）. The remaining mice were divided into sham operation group （n=20） and PSD group （n=18）. After injectied with NXN over-expression adeno-associated virus， the remaining mice in AAV-NXN-OE group and AAV-NC group were further divided into PSD+AAV-NC group （n=18） and PSD+AAV-NXN-OE group （n=18）. Three weeks before surgery， NXN over-expression adeno-associated virus was injected into the mPFC region of brain tissue of the mice by stereotaxic method， and the expression of the virus in mPFC region of the mice was observed under microscope. Western blotting method was used to detect the expression levels of NXN protein in brain tissue of the mice in various groups； middle cerebral artery occlusion （MCAO） model was established by thread embolism method， followed one week post-surgery by three weeks of chronic unpredictable moderate stress （CUMS） combined with isolation feeding to construct the PSD mice model. During modeling， the body weight changes of the mice were monitored. After modeling， sucrose preference test， tail suspension test， and forced swim test were used to observe the depressive-like behavioral changes of the mice in various groups； biochemical method was used to detect the levels of malondialdehyde （MDA） and reduced glutathione （GSH）， and superoxide dismutase （SOD） activities in mPFC region of brain tissue of the mice in various groups； DCFH-DA fluorescence probe labeling method was used to detect the reactive oxygen species （ROS） levels in mPFC region of brain tissue of the mice in various groups； Western blotting method was used to detect expression levels of NXN protein in mPFC region， amygdala， and hippocampus tissues of the mice in various groups. Results A large amount of green fluorescence was observed in the mPFC region in brain tissue of the PSD mice， indicating successful infection and expression of AAVs virus labeled with ZsGreen green fluorescent protein in mPFC region in brain tissue of the PSD mice. Compared with AAV-NC group， the expression level of NXN protein in mPFC region in brain tissue of the mice in AAV-NXN-OE group was significantly increased （P<0.05）. Compared with sham operation group， the body weight of the mice in PSD group was increased slowly （P<0.05）， the sucrose preference rate was significantly decreased （P<0.05）， and the immobility time of the mice in the tail suspension test and forced swim test was significantly increased （P<0.05）. Compared with sham operation group， the sucrose preference rate of the mice in PSD+AAV-NC group was significantly decreased （P<0.05）， and the immobility time in tail suspension test and forced swim test was significantly increased （P<0.05）. Compared with PSD+AAV-NC group， the sucrose preference rate of the mice in PSD+AAV-NXN-OE group was significantly increased （P<0.05）， and the immobility time of the mice in tail suspension test and forced swim test was significantly decreased （P<0.05）. Compared with sham operation group， the MDA and ROS levels in mPFC region in brain tissue of the mice in PSD+AAV-NC group were significantly increased （P<0.05）， while the GSH level and SOD activity were significantly decreased （P<0.05）. Compared with PSD+AAV-NC group， the levels of MDA and ROS in mPFC region in brain tissue of the mice in PSD+AAV-NXN-OE group were significantly decreased （P<0.05）， while the GSH level and SOD activity were significantly increased （P<0.05）. Compared with sham operation group， the expression level of NXN protein in the mPFC region of brain tissue of the mice in PSD group was significantly decreased （P<0.05）， the expression levels of NXN protein in amygdala and hippocampus tissue had no statistically significant difference （P>0.05）. Conclusion Over-expression of NXN in mPFC region of brain tissue of the mice can improve the depressive-like behavior in the PSD mice， and its mechanism is possibly related to regulating the redox balance.

Objective To discuss the protective effect of sodium butyrate（NaB） on acute liver injury in the mice induced by lipopolysaccharide（LPS） combined with D-galactosamine（D-Gal）， and to clarify its mechanism. Methods Thirty male Kunming mice were randomly divided into control group， model group， and NaB group， and there were 10 mice in each group. The mice in NaB group were given 200 mg·kg^-1·d^-1 NaB， while the mice in control group and model group were given an equal volume of sterile water. The mice in model group and NaB group were intraperitoneally injected with 20 μg·kg^-1 LPS and 600 mg·kg^-1 D-Gal to induce the acute liver injury models. The body weights and liver weights of the mice in various groups were detcted， and the liver index was calculated. HE staining was used to observe the pathomorphology of liver tissue of the mice in various groups； kits were used to detect the activities of alanine aminotransferase（ALT） and aspartate aminotransferase（AST） in serum， and the activities of total superoxide dismutase（T-SOD） and catalase（CAT）， and the levels of malondialdehyde（MDA） in liver tissue of the mice in various groups； Western blotting method was used to detect the expression levels of nuclear factor E2-related factor 2（Nrf2） and heme oxygenase-1（HO-1） proteins in liver tissue of the mice in various groups. Results There were no significant differences in body weights of the mice among various groups （P>0.05）. Compared with control group， the liver index of the mice in model group was significantly increased （P<0.01）. Compared with model group， the liver index of the mice in NaB group was significantly decreased （P<0.01）. The HE staining results showed that the liver tissue of the mice in control group exhibited normal structure， with clear boundaries of hepatocytes， consistent size， radially arranged around the central vein， and the nucleus located in the center of the cells； in model group， the arrangement of hepatocytes was disordered， the cells were swollen， there were multiple foci of hepatocellular necrosis， inflammatory cell infiltration， and hemorrhage； compared with model group， the cells in NaB group showed improved hepatocellular structure and reduced inflammatory infiltration. Compared with control group， the activities of ALT and AST in serum of the mice in model group were significantly increased （P<0.01）； compared with model group， the activities of ALT and AST in serum of the mice in NaB group were significantly decreased （P<0.05 or P<0.01）. Compared with control group， the activities of T-SOD and CAT in liver tissue of the mice in model group were significantly decreased （P<0.01）， and the level of MDA was significantly increased（P<0.01）； compared with model group， the activities of T-SOD and CAT in liver tissue of the mice in NaB group were significantly increased （P<0.05 or P<0.01）， and the level of MDA was significantly decreased（P<0.01）. The Western blotting results showed that compared with control group， the expression levels of Nrf2 and HO-1 proteins in liver tissue of the mice in model group were significantly decreased （P<0.05）； compared with model group， the expression levels of Nrf2 and HO-1 proteins in liver tissue of the mice in NaB group were significantly increased （P<0.01）. Conclusion NaB has a protective effect on LPS/D-Gal induced acute liver injury in the mice， and its mechanism may be related to the upregulation of the expressions of Nrf2 and HO-1 proteins and the increas of the activity of oxidant enzyme in liver tissue by NaB， thereby reduces the liver oxidative stress level of liver.

Objective To discuss the effect of parthenolide （PTL） on the apoptosis of the chondrocytes under mechanical stretch stress by regulating the expression of piezo type mechanosensitive ion channel component 1（Piezo1）， and to clarify the related mechanism. Methods The chondrocytes were divided into 0%， 5%， 10%， 15%， and 20% stretch groups according to the stretch variable. Additionally， the chondrocytes were divided into control group， 20% stretch group， 20% stretch+5 μmol·L^-1 PTL group， 20% stretch+10 μmol·L^-1 PTL group， and 20% stretch+20 μmol·L^-1 PTL group. The Piezo1 short hairpin RNA （shRNA） interference lentivirus （sh-Piezo1） or shRNA-NC lentivirus were used to infect the chondrocytes， and the chondrocytes were divided into sh-Piezo1 group and sh-NC group， and also set up blank control group. The chondrocytes were also devided into 20% stretch group， 20% stretch+PTL group， 20% stretch+sh-Piezo1 group， and 20% stretch+sh-Piezo1+PTL group. Hoechst 33258 fluorescence staining was used to observe the morphology of the nuclear in various groups； flow cytometry was used to detect the apoptotic rates of the cells in various groups； spectrophotometry was used to detect the cysteinyl aspartate specific proteinase（Caspase）-3 activities in the cells in various groups； CCK-8 method was used to detect the proliferation rates of the cells in various groups； Fluo-4/AM fluorescent probe method was used to detect the calicium ion（Ca²⁺） levels in the cells in various groups； real-time fluorescence quantitative PCR（RT-qPCR） method was used to detect the expression levels of Piezo1 mRNA in the cells in various groups； Western blotting method was used to detect the expression levels of Piezo1 protein in the cells in various groups. Results The Hoechst 33258 fluorescence staining resuts showed that as the increasing of stretch， the number of the chondrocytes with fragmented and densely stained nuclei in 0%， 5%， 10%， 15%， and 20% stretch groups were gradually increased. The flow cytometry results showed that compared with 0% stretch group， the apoptotic rates of the chondrocytes in 5%， 10%， 15%， and 20% stretch groups were significantly increased （P<0.01）； compared with control group， the apoptotic rate of the chondrocytes in 20% stretch group was significantly increased （P<0.05）； compared with 20% stretch group， the apoptotic rates of the chondrocytes in 20% stretch+ 5 μmol·L^-1 PTL group， 20% stretch+10 μmol·L^-1 PTL group， and 20% stretch+20 μmol·L^-1 PTL group were significantly decreased （P<0.05）； compared with 20% stretch group， the apoptotic rates of chondrocytes in 20% stretch+PTL group and 20% stretch+sh-Piezo1 group were significantly decreased （P<0.05）. The spectrophotometry results showed that compared with 0% stretch group， the Caspase-3 activities in the chondrocytes in 5%， 10%， 15%， and 20% stretch groups were significantly increased （P<0.01）； compared with control group， the Caspase-3 activity in the chondrocytes in 20% stretch group was significantly increased （P<0.05）； compared with 20% stretch group， the Caspase-3 activities in the chondrocytes in 20% stretch+5 μmol·L^-1 PTL group， 20% stretch+10 μmol·L^-1 PTL group， and 20% stretch+20 μmol·L^-1 PTL group were significantly decreased （P<0.05）. Compared with 20% stretch group， the Caspase-3 activities in the chondrocytes in 20% stretch+PTL group and 20% stretch+ sh-Piezo1 group were significantly decreased （P<0.05）. The CCK-8 method results showed that compared with 0 μmol·L^-1 PTL group， the proliferation rates of the chondrocytes in 40.00， 80.00， and 160.00 μmol·L^-1 PTL groups were significantly decreased （P<0.05）， indicating that 20.00 μmol·L^-1 PTL was the maximum non-toxic concentration. The Fluo-4/AM fluorescent probe method results showed that compared with control group， the Ca²⁺ level in the chondrocytes in 20% stretch group was significantly increased （P<0.05）； compared with 20% stretch group， the Ca²⁺ levels in the chondrocytes in 20% stretch+5 μmol·L^-1 PTL group， 20% stretch+10 μmol·L^-1 PTL group， and 20% stretch+ 20 μmol·L^-1 PTL group were significantly decreased （P<0.05）； compared with 20% stretch group， the Ca²⁺ levels in the chondrocytes in 20% stretch+PTL group and 20% stretch+sh-Piezo1 group were significantly decreased （P<0.05）. The RT-qPCR results showed that compared with blank control group and sh-NC group， the expression level of Piezo1 mRNA in the chondrocytes in sh-Piezo1 group was significantly decreased （P<0.05）. The Western blotting results showed that compared with control group， the expression levels of Piezo1 protein in the chondrocytes in 20% stretch group was significantly increased （P<0.05）； compared with 20% stretch group， the expression levels of Piezo1 protein in the chondrocytes in 20% stretch+5 μmol·L^-1 PTL group， 20% stretch+10 μmol·L^-1 PTL group， and 20% stretch+20 μmol·L^-1 PTL group were significantly decreased （P<0.05）； compared with blank control group and sh-NC group， the expression level of Piezo1 protein in the chondrocytes in sh-Piezo1 group was significantly decreased （P<0.05）. Conclusion PTL can inhibit the apoptosis of the chondrocyte induced by high-intensity cyclic mechanical stretch stress， and its mechanism may be related to inhibiting the Piezo1-mediated Ca²⁺ influx-induced apoptosis.

Objective To discuss the effect of microRNA （miR）-30c-5p on the proliferation， migration， and invasion of the human prostate cancer cells （LNCap）， and to clarify its possible mechanism. Methods The LNCap cells were divided into LNCap group （without plasmid transfection）， miR-30c-5p mimic group （transfected with miR-30c-5p mimic）， mimic NC group （transfected with miR-30c-5p mimic NC）， sh-DNA damage inducible transcript 4 （DDIT4） group （transfected with sh-DDIT4）， sh-NC group （transfected with sh-DDIT4 NC）， miR-30c-5p mimic+pc-DNA3.1-NC group （co-transfected with miR-30c-5p mimic and pc-DNA3.1 empty vector）， and miR-30c-5p mimic+pc-DNA3.1-DDIT4 group （co-transfected with miR-30c-5p mimic and pc-DNA3.1-DDIT4 over-expression plasmid）. The RWPE-1 cells were cultured normally.Real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of miR-30c-5p and DDIT4 mRNA in the cells in various groups； Western blotting method was used to detect the expression levels of DDIT4 protein in the cells in various groups； CCK-8 method was used to detect the proliferation rates of the LNCap cells in various groups； Transwell assay was used to detect the numbers of the invasion LNCap cells in various groups； Scratch assay was used to detect the scratch healing rates of LNCap cells in various groups； dual-luciferase reporter assay was used to detect the targeting relationship between miR-30c-5p and DDIT4.In the in vivo tumor formation experiment， 18 male BALB/c nude mice were divided randomly into blank group， agomiR-NC group （transfected with agomiR-30c-5p NC）， and agomiR-30c-5p group （transfected with agomiR-30c-5p）； there were six mice in each group.The mice in agomiR-NC group and agomiR-30c-5p group were subcutaneously injected with LNCap cells， while the mice in blank group were given an equal volume of physiological saline. The volumes of tumor of the mice in various groups were detected. HE staining was used to observe the morphology of prostate cancer tissue the mice of in various groups； RT-qPCR method and immunofluorescence staining were used to detect the expression levels of miR-30c-5p and DDIT4 mRNA and the fluorescence intensities of DDIT4 protein in prostate cancer tissue of the mice in various groups. Results The In vitro prostate cancer cell experiment results showed that compared with RWPE-1 cells， the expression level of miR-30c-5p in the prostate cancer LNCap cells was decreased （P<0.01）， and the expression levels of DDIT4 mRNA and protein were increased （P<0.05 or P<0.01）. After 48 of transfection， compared with LNCap group and mimic NC group， the expression level of miR-30c-5p in the LNCap cells in miR-30c-5p mimic group was increased （P<0.01）. Compared with LNCap group and sh-NC group， the expression level of DDIT4 mRNA in the LNCap cells in sh-DDIT4 group was decreased （P<0.01）. Compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group， the expression level of miR-30c-5p in The LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was decreased （P<0.01）； compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group， the expression level of DDIT4 mRNA in the LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was increased （P<0.01）； compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group， the expression level of DDIT4 protein in the LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was increased （P<0.05）. The CCK-8 method results showed that compared with LNCap group and mimic NC group， the proliferation rate of the LNCap cells in miR-30c-5p mimic group was decreased （P<0.01）； compared with LNCap group and sh-NC group， the proliferation rate of the LNCap cells in sh-DDIT4 group was decreased （P<0.01）； compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group， the proliferation rate of the LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was increased （P<0.01）. The Transwell assay results showed that compared with LNCap group and mimic NC group， the number of the invasion LNCap cells in miR-30c-5p mimic group was decreased （P<0.01）； compared with LNCap group and sh-NC group， the number of invasion LNCap cells in sh-DDIT4 group was decreased （P<0.01）； compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group， the number of the invasion LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was increased （P<0.01）.The scratch assay results showed that compared with LNCap group and mimic NC group， the scratch healing rate of the LNCap cells in miR-30c-5p mimic group was decreased （P<0.01）； compared with LNCap group and sh-NC group， the scratch healing rate of the LNCap cells in sh-DDIT4 group was decreased （P<0.01）； compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group， the scratch healing rate of the LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was increased （P<0.01）. The dual-luciferase reporter assay results showed that compared with the LNCap cells co-transfected with WT-DDIT4 and mimic NC， the luciferase activity of the LNCap cells co-transfected with WT-DDIT4 and miR-30c-5p mimic was decreased （P<0.01）. The in vivo nude mouse tumor formation experiment results showed that on the 3 rd， 6 th， 9 th， 12 th， and 15th days after cell injection， compared with blank group and agomiR-NC group， the tumor volumes of the nude mice in agomiR-30c-5p group were decreased （P<0.05）. The HE staining results showed that in prostate cancer tissue of the mice in blank group and agomiR-NC group， the cell nuclei were enlarged， and nucleoli were prominent and deformed. In the mice in agomiR-30c-5p group， some regions of prostate cancer tissues results showed neatly arranged cells with normally shaped nuclei. The RT-qPCR and immunofluorescence staining showed that compared with agomiR-NC group， the expression level of miR-30c-5p in prostate cancer tissue of the mice in agomiR-30c-5p group was increased （P<0.01）. Compared with blank group and agomiR-NC group， the expression level of DDIT4 mRNA in prostate cancer tissue of the mice in agomiR-30c-5p group was decreased （P<0.01）. DDIT4 protein was mainly expressed in the cytoplasm. Compared with blank group and agomiR-NC group， the fluorescence intensity of DDIT4 protein in prostate cancer tissue of the mice in agomiR-30c-5p group was decreased （P<0.01）. Conclusion The expression level of miR-30c-5p in the prostate cancer LNCap cells is decreased， and it inhibits the proliferation， migration， and invasion of the prostate cancer cells by targeting downregulation of DDIT4， thereby participating in the occurrence and development of prostate cancer.

Objective To discuss the effect of knockdown of receptor-interacting protein kinase 3 （RIP3） on autophagy， pyroptosis， and ferroptosis in the human renal tubular epithelial HK2 cells under hypoxia/reoxygenation （H/R） conditions. Methods The lentiviral interference vector plasmid shRIP3 and negative control lentiviral interference vector plasmid shNC were transfected into the HK2 cells and the HK cells were divided into shRIP3 group and shNC group， and the normal cultured untransfected HK2 cells were regarded as blank group. After 48 h of transfection， real-time fluorescence quantitative PCR （RT-qPCR） and Western blotting methods were used to verify the lentiviral transfection efficiencies. The HK2 cells were divided into control group， H/R group， shNC+H/R group， and shRIP3+H/R group. CCK-8 method was used to detect the survival rates of the HK2 cells in various groups； immunofluorescence staining was used to detect the fluorescence intensities of light chain 3 （LC3B） and NLR family pyrin domain-containing 3 （NLRP3） proteins in the cells in various groups； Western blotting method was used to detect the expression levels of LC3Ⅱ， LC3Ⅰ， Beclin1， Caspase-1， Gasdermin D （GSDMD）， interleukin （IL）-1β， and IL-18 proteins in the HK2 cells in various groups； Kits were used to detect the ferri ion（Fe²⁺） levels in the cells in various groups. Results Compared with blank group and shNC group， the expression levels of RIP3 mRNA and protein in the HK2 cells in shRIP3 group were decreased （P<0.05）.The CCK-8 method results showed that compared with control group， the survival rate of the HK2 cells in H/R group was decreased （P<0.05）； compared with H/R group， the survival rate of the HK2 cells in shRIP3+H/R group was increased （P<0.05）. The immunofluorescence staining results showed that compared with control group， the fluorescence intensity of LC3B protein in the HK2 cells in H/R group was decreased （P<0.05）， and the fluorescence intensity of NLRP3 protein was increased （P<0.05）； compared with H/R group， the fluorescence intensity of LC3B protein in the HK2 cells in shRIP3+H/R group was increased （P<0.05）， and the fluorescence intensity of NLRP3 protein was decreased （P<0.05）. The Western blotting results showed that compared with control group， the ratio of LC3Ⅱ/LC3Ⅰ and the expression level of Beclin1 protein in the HK2 cells in H/R group were decreased （P<0.05）； compared with H/R group， the ratio of LC3Ⅱ/LC3Ⅰ and expression level of Beclin1 protein in the HK2 cells in shRIP3+H/R group were increased （P<0.05）； compared with control group， the expression levels of Caspase-1， GSDMD， IL-1β， and IL-18 proteins in the HK2 cells in H/R group were increased （P<0.05）； compared with H/R group， the expression levels of Caspase-1， GSDMD， IL-1β， and IL-18 proteins in the HK2 cells in shRIP3+H/R group were decreased （P<0.05）. Compared with control group， the expression levels of GPX4 and SLC7A11 proteins in the HK2 cells in H/R group were decreased （P<0.05）； compared with H/R group， the expression levels of GPX4 and SLC7A11 proteins in the HK2 cells in shRIP3+H/R group were increased （P<0.05）. Compared with control group， the Fe²⁺ level in the HK2 cells in H/R group was increased （P<0.05）； compared with H/R group， the Fe²⁺ level in the HK2 cells in shRIP3+H/R group was decreased （P<0.05）. Conclusion Targeted knockdown of RIP3 can induce the autophagy， inhibit the pyroptosis， and reduce the ferroptosis of the human renal tubular epithelial HK2 cells induced by H/R.

Objective To discuss the effect of Fuzheng Ruanjian Anticancer Formula on the malignant biological behaviors of the hepatocellular carcinoma HepG2 cells by requlating protein kinase B（Akt）/murine double minute 2（MDM2）/P53 signaling pathway. Methods The HepG2 cells were treated with 0， 0.05， 0.10， 0.20， 0.40， 0.80， 1.60， 3.20， and 6.40 g·mL^-1 Fuzheng Ruanjian Anticancer Formula for 48 h. CCK-8 method was used to detect the survival rates of the HepG2 cells in various groups， and the concentrations of Fuzheng Ruanjian Anticancer Formula for the subsequent experiments were screened. The HepG2 cells were divided into control group， low dose of Fuzheng Ruanjian Anticancer Formula group （0.2 g·mL^-1）， medium dose of Fuzheng Ruanjian Anticancer Formula group （0.4 g·mL^-1）， high dose of Fuzheng Ruanjian Anticancer Formula group （0.8 g·mL^-1）， SC79 group （8 mg·L^-1 SC79）， and high dose of Fuzheng Ruanjian Anticancer Formula+SC79 group （0.8 g·mL^-1 Fuzheng Ruijian Anticancer Formula+8 mg·L^-1 SC79）. CCK-8 method was used to detect the proliferation activities of the HepG2 cells in various groups； clone formation assay was used to detect the clone formation rates of the HepG2 cells in various groups； flow cytometry was used to detect the apoptotic rates of the HepG2 cells in various groups； Transwell chamber assay was used to detect the numbers of migration and invasion HepG2 cells in various groups； Western blotting method was used to detect the expression levels of proliferating cell nuclear antigen （PCNA）， cysteine aspartate specific proteinase （Caspase-3）， matrix metalloproteinase （MMP）-2， MMP-9， phosphorylated Akt （p-Akt）， phosphorylated MDM2 （p-MDM2）， and P53 proteins in the HepG2 cells in various groups. Results As the increasing of concentrations of Fuzheng Ruanjian Anticancer Formula （0， 0.05， 0.10， 0.20， 0.40， 0.80， 1.60， 3.20， and 6.40 g·mL^-1）， the surival rates of the HepG2 cells were gradually decreased （P<0.05）， and 0.2， 0.4， and 0.8 g·mL^-1 Fuzheng Ruanjian Anticancer Formula were selected for the subsequent experiments. The CCK-8 assay results showed that compared with control group， the proliferation activities of the HepG2 cells in low， medium， and high doses of Fuzheng Ruanjian Anticancer Formula groups were significantly decreased （P<0.05）， in a dose-dependent manner， while the proliferation activity of the cells in SC79 group was significantly increased （P<0.05）. Compared with high dose of Fuzheng Ruanjian Anticancer Formula group， the proliferation activity of the HepG2 cells in high dose of Fuzheng Ruanjian Anticancer Formula+SC79 group was significantly increased （P<0.05）. The clone formation assay results showed that compared with control group， the clone formation rates of the HepG2 cells in low， medium， and high doses of Fuzheng Ruanjian Anticancer Formula groups were significantly decreased （P<0.05） in a dose-dependent manner， while the clone formation rate of the cells in SC79 group was significantly increased （P<0.05）； compared with high dose of Fuzheng Ruanjian Anticancer Formula group， the clone formation rate of the cells in high dose of Fuzheng Ruanjian Anticancer Formula+SC79 group was significantly increased （P<0.05）. The flow cytometry results showed that compared with control group， the apoptotic rates of the HepG2 cells in low， medium， and high doses of Fuzheng Ruijian Anticancer Formula groups were significantly increased （P<0.05） in a dose-dependent manner， while the apoptotic rate of the cells in SC79 group was significantly decreased （P<0.05）； compared with high dose of Fuzheng Ruanjian Anticancer Formula group， the apoptotic rate of the cells in high dose of Fuzheng Ruanjian Anticancer Formula+SC79 group was significantly decreased（P<0.05）. The Transwell chamber assay results showed that compared with control group， the numbers of migration and invasion HepG2 cells in low， medium， and high doses of Fuzheng Ruanjian Anticancer Formula groups were significantly decreased （P<0.05） in a dose-dependent manner， while the numbers of migration and invasion cells in SC79 group were significantly increased （P<0.05）； compared with high dose of Fuzheng Ruanjian Anticancer Formula group， the numbers of migration and invasion HepG2 cells in high dose of Fuzheng Ruanjian Anticancer Formula+SC79 group were significantly increased （P<0.05）. The Western blotting results showed that compared with control group， the expression levels of PCNA， MMP-2， MMP-9， p-Akt， and p-MDM2 proteins in the cells in low， medium， and high doses of Fuzheng Ruanjian Anticancer Formula groups were significantly decreased （P<0.05） in a dose-dependent manner， while the expression levels of Caspase-3 and P53 proteins were significantly increased （P<0.05） in a dose-dependent manner， while the expression levels of PCNA， MMP-2， MMP-9， p-Akt， and p-MDM2 proteins in the cells in SC79 group were significantly increased （P<0.05）， and the expression levels of Caspase-3 and P53 proteins were significantly decreased （P<0.05）； compared with high dose of Fuzheng Ruanjian Anticancer Formula group， the expression levels of PCNA， MMP-2， MMP-9， p-Akt， and p-MDM2 proteins in the cells in high dose of Fuzheng Ruanjian Anticancer Formula+SC79 group were significantly increased （P<0.05）， while the expression levels of Caspase-3 and P53 proteins were significantly decreased （P<0.05）. Conclusion Fuzheng Ruanjian Anticancer Formula may inhibit the proliferation， migration， and invasion of the HepG2 cells and promote the apoptosis， and its mechanism may be related to suppressing the Akt/MDM2 signaling pathway and upregulating the P53 proteim expression.

Objective To screen for regulatory cell death and senescence genes with differential prognostic significances in the gastric cancer through bioinformatics methods， and to analyze the effect of phosphodiesterase 1B （PDE1B） on the survival prognosis of the gastric cancer patients. Methods The gastric cancer gene expression data and clinical data were downloaded from the TCGA Public Database.Fifty gastric cancer patients were randomly selected from the local database， and their clinical informations and paraffin samples， including gastric cancer tissue and adjacent normal tissue， were collected. The R software “limma” package was used to screen differentially expressed genes （DEGs）； univariate COX analysis and Kaplan-Meier survival analysis were used to screen DEGs with predictive survival value. The intersection genes affecting the survival prognosis of gastric cancer patients were obtained， and the gene most associated with clinical pathological features PDE1B was screened. The TCGA Database and Kaplan-Meier survival analysis were used to detect the expression levels of PDE1B mRNA in adjacent normal and gastric cancer tissues and their relationships with survival period of the gastric cancer patients. Gene Ontology （GO） functional and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analyses were used to enrich the biological functions of PDE1B. The CIBERSORT algorithm， Tumor Immunity Database （TISIDB）， and GSCA online website were used to analyze the correlation between PDE1B and tumor microenvironment， immune characteristic molecules， and drug sensitivity. The real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the PDE1B mRNA expression levels in gastric cancer tissue and adjacent normal tissue of the gastric cancer patients. Results A total of 716 DEGs were screened， among which 505 DEGs were upregulated （P<0.05）， and 211 DEGs were downregulated （P<0.05）. There were 10 intersection genes affecting survival prognosis.The PDE1B mRNA expression level was most closely related to the clinical pathological characteristics of the gastric cancer patients， being associated with age， tumor grade， tumor stage， and tumor T， N， and M stages （P<0.05）. Compared with G1-G2， StageⅠ， T1-T2， N0， and M0 stage gastric cancer patients，the PDE1B mRNA expression levels in G3-G4， StageⅡ-Ⅳ， T3-T4， N1-N3， and M1 stage gastric cancer patients were significantly increased （P<0.05）. Compared with adjacent normal tissue， the PDE1B mRNA expression level in gastric cancer tissue was significantly decreased （P<0.05）. Compared with the patients with low PDE1B expression， the patients with high PDE1B expression had a significantly lower overall survival rate （P<0.01）. PDE1B expression， age， and tumor stage were the risk factors for the prognosis of gastric cancer patients （P<0.05）. After adjusting for gender， age， tumor grade， and tumor stage， PDE1B expression was an independent risk factor affecting the prognosis of the gastric cancer patients （P<0.05）. PDE1B was mainly enriched in the biological process（BP）， such as immunoglobilin production， second-messenger， mediated signaling transduction and calcium ion transport， cellular component（CC）， such as Tlymplocytes receptor complex， plasma membrance signaling receptor complex and collagen-containing extracellular matrix， and molecular function（MF）， such as antigen binding， glycosaminoglycan binding， and extracellular matrix structural constituents. PDE1B was mainly involved in the pathways such as neuroactive ligand-receptor interaction， calcium signaling pathway， cGMP-PKG signaling pathway， and cytokine-cytokine receptor interaction. PDE1B was positively correlated with regulatory T lymphocytes （r=0.488）， myeloid-derived suppressor cells （r=0.474）， and macrophages （r=0.617） （P<0.01）. Compared with the patients with low PDE1B expression， the patients with high PDE1B expression promoted the significant increase of infiltration of regulatory T lymphocytes， monocytes， and M2 macrophages （P<0.05）. The PDE1B mRNA expression levels were positively correlated with the immunosuppressive agents transforming growth factor-β1 （TGF-β1） （r=0.535）， colony-stimulating factor 1 receptor （CSF1R） （r=0.519）， immune activator ectonucleoside triphosphate diphosphohydrolase 1 （ENTPD1） （r=0.593）， and CXC chemokine ligand 12 （CXCL12） （r=0.646） （P<0.01）. The gastric cancer tissue with high PDE1B expression was more sensitive to the drugs such as fluorouracil （-0.3<r <-0.1）， methotrexate （-0.3<r <-0.1）， and decitabine （-0.3<r <-0.1） （P<0.05）. Compared with adjacent normal tissue， the PDE1B mRNA expression levels in gastric cancer tissue were significantly decreased （P<0.01）. Compared with low PDE1B expression group， the overall survival rate patients in of the high PDE1B expression group had a significantly lower was decreased（P<0.05）.Compared with T1-T2 stage gastric cancer patients， the PDE1B mRNA expression level of the T3-T4 stage gastric cancer patients was significantly increased （P<0.01）. Conclusion PDE1B is an independent risk factor for the prognosis of the gastric cancer patients and can serve as an effective indicator of poor prognosis of gastric cancer.

Objective To discuss the clinical characteristics of the patients with ovarian clear cell carcinoma （OCCC） and ovarian endometriosis （OMA）， and to clarify the features of OCCC onset. Methods A retrospective analysis was conducted on the clinical data of 80 patients with post-operative pathological diagnosis of OCCC and 80 OMA patients diagnosed by post-operative pathology， who received surgical treatment from March 2011 to May 2021. The analysis included general characteristics， clinical manifestations， laboratory indices， and imaging examination indexes. The age， body mass index （BMI）， clinical symptoms such as abdominal pain， vaginal bleeding， and other symptoms （bloating， menstrual disorders， constipation， and abnormal vaginal discharge）， preoperative serum carbohydrate antigen 125 （CA125） levels， and ultrasound characteristics of ovarian cysts， such as cyst size， presence of pelvic effusion， and complexity， were compared between the patients in two groups. Multivariate Logistic regression analysis was used to identify the risk factors for OCCC， and the receiver operating characteristic （ROC） curve was drawn， and the area under the curve （AUC） was calculated. Results Compared with OMA group， the age of the patients in OCCC group was significantly increased （P<0.05）， the serum CA125 level was significantly increased （P<0.05）， the diameter of ovarian cyst was significantly increased （P<0.05）， and the percentage of the patients with pelvic effusion， other symptoms， and complex cysts was significantly increased （P<0.05）. The multivariate Logistic regression analysis results showed that age≥40 years （OR=56.856， 95%CI： 5.611-576.082， P=0.001）， complex ovarian cysts （OR=4.427， 95%CI： 1.025-19.114， P=0.046）， concomitant pelvic effusion （OR=8.760， 95%CI： 1.574-48.760， P=0.013）， and size of ovarian cysts （OR=1.782， 95%CI： 1.329-2.390， P<0.01） were the risk factors for OCCC. When the cut-off value of the ovarian cyst size was 7.35 cm， the sum of sensitivity （83.75%） and specificity （80.00%） was the highest， and the AUC was 0.883， indicating certain predictive value for identifying OCCC. Conclusion The patients aged≥40 years with cysts≥7.35 cm in diameter， especially those with complex cysts accompanied by pelvic effusion， have a higher risk of malignancy to OCCC， and active intervention is recommended.

Objective To evaluate the clinical efficacy and safety of secukinumab in the treatment of moderate to severe the adults with plaque psoriasis. Methods The clinical data from 183 adult patients with moderate to severe plaque psoriasis treated with secukinumab were collected. The patients received subcutaneous injections of secukinumab once a week at weeks 0， 1， 2， 3， and 4， followed by an injection every 4 weeks， with each dose of 300 mg. The follow-up period was 52 weeks. The psoriasis area and severity index（PASI）， body surface area （BSA）， investigator global assessment （IGA）， and dermatology life quality index （DLQI） scores of the patients with psoriasis were caculated. The clinical efficacy and safety of secukinumab in the treatment of moderate to severe plaque psoriasis were evaluated， and the influencing factors were analyzed. Results Compared with week 0， the PASI、 BSA、 IGA and DLQI scores of the patients were significantly decreased at weeks 4， 12， 24， and 52 of secukinumab treatment （P<0.05）. After treated with secukinumab， the percentages of the patients achieving PASI 75， PASI 90， and PASI 100 at week 4 were 95.6%， 84.2%， and 47.5%， respectively； at week 12， they were 97.3%， 95.6%， and 78.7%， respectively； at week 24， they were 97.8%， 96.7%， and 84.2%， respectively； and at week 52， they were 98.4%， 97.8%， and 83.6%， respectively. The percentages of the patients with BSA≤1% at weeks 4， 12， 24， and 52 were 80.9%， 94.5%， 95.6%， and 94.0%， respectively. The percentages of the patients with IGA score of 0/1 at week 4， 12， 24， and 52 were 86.3%， 97.3%， 96.7%， and 95.6%， respectively. The percentages of the patients with DLQI score of 0/1 at weeks 4， 12， 24， and 52 were 76.6%， 89.1%， 92.9%， and 91.8%， respectively. At week 4 of secukinumab treatment， there were significant differences in age， body mass index （BMI）， disease duration， baseline PASI score， and history of previous biologic treatment between the patients in two groups （P<0.05）. At week 24 of secukinumab treatment， there were significant differences in age and BMI between the patients in two groups （P<0.05）. At week 4， BMI≥25 kg·m^-2， disease duration≥10 years， baseline PASI score≥10， and a history of previous biologic treatment were risk factors affecting the recovery of the patient （P<0.05）. At week 24， age≥40 years was a risk factor affecting the recovery of the patient （P<0.05）. During the treatment period， 44 out of 183 psoriasis patients reported 49 adverse reactions， and the adverse reaction rate was 24.0%. No serious adverse events or fatal adverse reactions occurred. The adverse reactions included upper respiratory tract infections in 23 cases， eczema-like skin lesions in 10 cases， skin fungal infections in 6 cases， urticaria in 3 cases， mild liver function abnormalities in 2 cases， folliculitis in 2 cases， conjunctivitis in 2 cases， and otitis media in 1 case. Conclusion Secukinumab treatment for the adult patients with moderate to severe plaque psoriasis is rapid-acting and has lasting efficacy. The BMI， disease duration， baseline PASI score， history of previous biologic treatment， and age are the factors influencing the clinical efficacy of secukinumab. The overall safety is good， and secukinumab may be used as a first-line treatment option for moderate to severe plaque psoriasis.

Objective To discuss the differences in eosinophil (EOS) infiltration in cervical tissue and its relationship with cervical-related diseases, and to clarify the effect of EOS on the occurrence and development of cervical intraepithelial neoplasia (CIN) and cervical cancer. Methods The clinical data of 256 patients with cervical diseases were collected and divided into cervical cancer group （n=46, including 26 cases of squamous cell carcinoma, 15 cases of adenocarcinoma, and 5 cases of adenosquamous carcinoma), chronic cervicitis group (n=50), CIN stageⅠ group (n=50), CIN stageⅡ group (n=50), CIN stageⅢ group (n=30), and normal group (adjacent normal cervical tissue, n=30) based on their conditions. Colposcopy was used to observe the morphology of cervical tissue of the patients in various groups； thin-layer liquid-based cytology test (TCT) was used to observe the morphology of the cervical exfoliated cells in various groups； hybrid capture-chemiluminescence method was used to detect the human papillomavirus (HPV) infection in cervical tissue of the patients in various groups； HE staining was used to observe the pathomorphology of cervical tissue of the patients in various groups； Congo red staining was used to detect the numbers of EOS infiltration in cervical tissue of the patients in various groups； Pearson correlation analysis was used to analyze the correlation between the number of EOS infiltration and the malignancy degree of cervical cancer. Results The cervical surface of the patients in normal group was smooth and pink, with uniformly distributed capillaries； the cervical surface of the patients in chronic cervicitis group showed red inflammatory changes, with some accompanied by Nabothian cysts and varying degrees of erosion and ulcers； the patients in CIN stageⅠ, CIN stageⅡ, and CIN stageⅢ groups showed epithelial ulcers, thickening, and irregular morphology, with mosaic and punctate vessels； the cervical surface of the patients in cervical cancer group showed raised areas with neoplasms and necrotic ulcers, and they were fragile and prone to bleeding. After acetic acid staining, no obvious changes of the patients in normal group were observed. The cervix of the patients in chronic cervicitis group showed slight white changes that lasted for a short time； in CIN stageⅠ, CIN stageⅡ, and CIN stageⅢ groups, irregular thin acetowhite epithelium with map-like borders was observed, with increasingly acetowhite reactions and larger areas as the stages advanced. The cervix of the patients in cervical cancer group showed thick acetowhite epithelium that lasted longer, with rigid and clear contours. After iodine staining, the cervix of the patients in normal group was brown, with uniform coloration; the cervix of the patients in chronic cervicitis group showed poor coloration in inflammatory lesion areas； the cervix of the patients in CIN stageⅠ group showed iodine coloration in metaplastic areas, while the cervix of the patients in CIN stageⅢ group showed poor coloration in larger lesion areas； the cervix of the patients in cervical cancer group showed irregular surfaces with cauliflower-like growth and no coloration after iodine staining, appearing orange-yellow or mustard yellow. The TCT observation results showed there were no heteromorphic cells and few inflammatory cells in cervical exfoliated cells of the patients in infiltration in normal group； there were numerous neutrophils and EOS in exfoliated cervical cells without heteromorphic cells in chronic cervicitis group. The heteromorphic binucleated cells with high nuclear-cytoplasmic ratios and deeply stained nuclei were observed in cervical exfoliated cells of the patients in CIN stageⅠ and CIN stageⅡ groups. More heteromorphic cells with high nuclear-cytoplasmic ratios and irregular nuclear membranes were showed in cervical exfoliated cells of the patients in CIN stageⅢ group. The cervical exfoliated cells of the patients in cervical cancer group showed large and prominent nucleoli, clustering into syncytial changes. Compared with normal group, the atypial of cervical exfoliated cells in CIN stageⅠ, CIN stageⅡ, CIN stageⅢ, and cervical cancer groups was increased. The hybrid capture-chemiluminescence results showed that compared with normal and chronic cervicitis groups, the numbers of HPV infection and TCT heteromorphic cells of the patients in CIN stageⅠ, CIN stageⅡ, and CIN stageⅢ groups were increased (P<0.05)； compared with CIN stageⅠ, CIN stageⅡ, and CIN stageⅢ groups, the numbers of HPV infection and TCT heteromorphic cells of the patients in cervical cancer group were increased (P<0.05). The HE staining results showed normal cell morphology and structure in normal group, with infiltration of inflammation cells such as neutrophils, monocytes, macrophages, EOS, and lymphocytes； in chronic cervicitis group, the infiltration of inflammatory cells was increased； in CIN group, the cervical cells showed slightly larger nucleoli and heteromorphic cells, with inflammatory cells mainly distributing around the hetermomorphic cells； in cervical cancer group, the cervical cells showed large and deeply stained nucleoli with significant atypia, and the infiltration of inflammatory cells around the cancer cells was increased. Compared with normal group, the numbers of inflammatory cells and EOS infiltration in cervical tissue of the patients in chronic cervicitis group were increased (P<0.05), and the numbers of inflammatory cells and EOS infiltration of the patients in CIN group were increased (P<0.05)； compared with chronic cervicitis group, the number of inflammatory cells and EOS infiltration of the patients in CIN group were decreased (P<0.05)； compared with chronic cervicitis group and CIN group, the numbers of inflammatory cells and EOS infiltration of the patients in cervical cancer group were increased (P<0.05). The EOS in cervical cancer tissue was mainly distributed around the cancer nests； compared with CIN stageⅠ group, the numbers of EOS infiltration in CIN stageⅡ and CIN stageⅢ groups were increased （P<0.05）； compared with CIN stageⅡ group, the number of EOS infiltration in CIN stageⅢ group was increased （P<0.05）. The higher the malignancy degree of the tumor, the more EOS infiltration was observed, and the number of EOS infiltration was positively correlated with the invasion depth of cervical cancer （r=0.533 0, P<0.01）. Conclusion HPV infection and EOS infiltration play a role in promoting the and occurrence development of cervical precancerous lesions and cervical cancer.

Objective To discuss the clinical efficacy of recombinant human growth hormone （rhGH） in the treatment of the pediatric patients with growth hormone deficiency （GHD） and idiopathic short stature （ISS）， and to clarify its clinical application value in the pediatric patients with short stature of different etiologies. Methods The clinical data of 132 children with short stature who treated with rhGH from January 2018 to January 2023 were collected. They were divided into GHD group （n=70） and ISS group （n=62） based on different etiologies. The bone age， target height （TH）， body mass index （BMI）， height standard deviation score （HtSDS）， changes in height standard deviation scores（ΔHtSDS） before treatment and 6 months after treatment， and growth velocity （GV） of the pediatric patients were calculated. Propensity score matching （PSM） and inverse probability of treatment weighting （IPTW） were used to balance the confounding factors between the pediatric patients in two groups and the efficacy and safety of the pediatric patients in two groups were evaluated. Results There were significant differences in whether children were full-term， bone age， bone age maturity， and TH of the pediatric patients between two groups （P<0.05）. Compared with before treatment， the height and HtSDS of the pediatric patients in both GHD and ISS groups were significantly increased after treated for 6 months （P<0.05）. Before matched by PSM， there were significant differences in full-term， bone age， bone age maturity， and TH of the pediatric patients between two groups （P<0.05）. After matched by PSM， there were no significant differences in gender， region， term birth status， mode of delivery， feeding method， age， bone age， height， BMI， TH， and pretreatment HtSDS of the pediatric patients between two groups （P>0.05）； the standardized mean difference （SMD） differences of covariates except for region were<0.2. After weighted by IPTW，there were no significant differences in gender， region， term birth status， mode of delivery， feeding method， age， bone age， height， BMI， TH， and pretreatment HtSDS of the pediatric patients between two groups （P>0.05）； all SMD of covariates except for term birth status were<0.2. Before balancing covariates， after meatched by PSM matching， and after weighted by IPTW weighting compared with GHD group，the GV and ΔHtSDS of the pediatric patients in ISS group were slightly increased， but the difference was not significant （P>0.05）. In terms of adverse reactions， 2 cases （2.68%） of fasting hyperglycemia and 7 cases （10.00%） of hypothyroidism occurred in GHD group； 3 cases （4.84%） of fasting hyperglycemia and 2 cases （3.23%） of hypothyroidism occurred in ISS group. Conclusion rhGH can promote the height increase in the patients with GHD and ISS， and there is no significant difference in the height-increasing efficacy between GHD and ISS children. The incidence of adverse reactions is relatively low during treatment， indicating good overall safety.

Objective To analyze the association between the rs562556 polymorphism of the proprotein convertase subtilisin/kexin type 9（PCSK9） gene and the degree of coronary artery stenosis in the patients with acute myocardial infarction（AMI）. Methods A total of 200 patients diagnosed with AMI from January 2021 to December 2022 were selected as AMI group， and 200 healthy individuals during the same period were selected as control group. According to the Gensini scoring standard， the patients in AMI group were divided into low risk group （Gensini score≤40， n=78） and medium-high risk group（Gensini score>40， n=122）. The levels of lipid metabolism indicators in serum of the patients in two groups were detected by fully automatic biochemical analyzer； enzyme linked immunosorbent assay （ELISA） method was used to detect the PCSK9 levels in serum of the patients in two groups； ultraviolet spectrophotometry was used to detect the single nucleotide polymorphism （SNP） of PCSK9 gene of the patients in two groups； Spearman correlation analysis was used to detect the correlation between the rs562556 polymorphism of the PCSK9 gene and the degree of the disease and the levels of lipid metabolism indicators in serum of the patients. Results Compared with control group， the percentage of smokers of the patients in AMI group was significantly increased（P<0.01）. Compared with control group， the levels of low-density lipoprotein cholesterol （LDL-c） and PCSK9 in serum of the patients in AMI group were significantly increased（P<0.05）. Compared with low risk group， the levels of LDL-c and PCSK9 in serum of the patients in medium-high risk group were significantly increased（P<0.05）. The distribution of PCSK9 gene rs1800487 genotype in both control and AMI groups conformed to the Hardy-Weinberg（H-W） equilibrium（χ²=0.677， P=0.713； χ²=0.970， P=0.831）. Compared with control group， the distribution frequencies of PCSK9 gene rs562556 genotype AA and allele A of the patients in AMI group were significantly increased（P<0.05）. In the AMI patients， the distribution of PCSK9 gene rs562556 genotype in both low risk and medium-high risk groups conformed to the H-W equilibrium（χ²=0.045， P=0.978； χ²=1.290， P=0.525）. Compared with low risk group， the distribution frequencies of genotype AA and allele A of PCSK9 gene rs562556 of the patients in medium-high risk group were significantly increased（P<0.05）. The PCSK9 gene rs562556 genotype AA was positively correlated with the degree of AMI（r=0.193， P=0.006） and LDL-c level（r=0.301， P<0.01）. Allele A was positively correlated with the LDL-c level（r=0.168， P=0.017）. Conclusion The PCSK9 gene rs562556 genotype AA is positively correlated with the degree of coronary artery stenosis of the AMI patients， and its polymorphism may promote the development of AMI by upregulating the LDL-c level.

Objective To discuss the differences in serum proline dehydrogenase （ProDH） levels among chronic heart failure （CHF） patients with different ejection fraction types， and to clarify the effect of ProDH levels on cardiac function. Methods A retrospective analysis of clinical data of 118 CHF patients was conducted. These patients were divided into heart failure with reduced ejection fraction （HFrEF） group （n=39）， heart failure with mid-range ejection fraction group （HFmrEF） （n=42）， and heart failure with preserved ejection fraction （HFpEF） group （n=37）. A total of 45 non-CHF patients hospitalized during the same period were collected as control group. The general data of all the subjects in various groups were collected， and the levels of biochemical indicators and cardiac structure indicators in serum of all the subjects were detected. Spearman correlation analysis and point-biserial correlation analysis were used to analyze the correlation between serum ProDH levels and various biochemical indicators； multivariate Logistic regression analysis was used to analyze the factors influencing HFrEF and HFmrEF. Results Compared with control group， the usage rate of beta-blockers of the patients in HFpEF group was significantly increased （P<0.05）； in HFmrEF group， the percentage of male patients， the usage rate of statins， and the usage rate of beta-blockers were all significantly increased （P<0.05）； in HFrEF group， the age and systolic blood pressure （SBP） of the patients were significantly decreased （P<0.05）， while the usage rates of statins and beta-blockers of the patients were significantly increased （P<0.05）. Compared with HFpEF group， the age of the patients in HFmrEF group was significantly decreased （P<0.05）， and the percentage of male patients and the usage rate of statins were significantly increased （P<0.05）； the age of the patients in the HFrEF group was significantly decreased （P<0.05）， and the usage rate of statins was significantly increased （P<0.05）. Compared with HFmrEF group， the SBP of the patients in HFrEF group was significantly decreased （P<0.05）.Compared with control group， the serum levels of low-density lipoprotein cholesterol （LDL-c） of the patients in HFpEF and HFmrEF groups were significantly decreased （P<0.05）， while the levels of N-terminal pro-brain natriuretic peptide （NT-proBNP） were significantly increased （P<0.05）； the serum levels of glomerular filtration rate （GFR） and ProDH of the patients in HFrEF group were significantly decreased （P<0.05）， and the levels of fasting blood glucose （FBG） and NT-proBNP were significantly increased （P<0.05）. Compared with HFpEF group， the serum hemoglobin （Hb） level of the patients in HFmrEF group was significantly increased （P<0.05）； the serum NT-proBNP level of the patients in HFrEF group was significantly increased （P<0.05）， while the ProDH level was significantly decreased （P<0.05）. Compared with HFmrEF group， the serum NT-proBNP level of the patients in HFrEF group was significantly increased （P<0.05）.Compared with control group， the left atrial diameter （LAD） and the ratio of early diastolic mitral inflow velocity to early diastolic mitral annular velocity （E/Em） of the patients in HFpEF， HFmrEF， and HFrEF groups were significantly increased （P<0.05）； the left ventricular end-diastolic diameter （LVEDD） of the patients in HFmrEF and HFrEF groups were significantly increased （P<0.05）， and the left ventricular ejection fraction （LVEF） were significantly decreased （P<0.05）. Compared with HFpEF group， the LVEDD of the patients in HFmrEF and HFrEF groups were significantly increased （P<0.05）， and the LVEF were significantly decreased （P<0.05）； the LAD of the patients In HFrEF group was significantly increased （P<0.05）. Compared with HFmrEF group， the E/Em ratio of the patients in HFrEF group was significantly increased （P<0.05）， and the LVEF was significantly decreased（P<0.05）. The serum ProDH levels of the patients were negatively correlated with LVEDD （r=-0.210， P=0.007） and positively correlated with LVEF （r=0.220， P=0.005）. Male and elevated FBG levels were the risk factors for cardiac function， while the increasing serum GFR and ProDH levels were the protective factors for cardiac function. Conclusion There are differences in ProDH levels among the CHF patients with different ejection fraction types. The patients with poorer cardiac function have lower serum ProDH levels， and higher ProDH levels may be beneficial for improving the left ventricular systolic function in the CHF patients.KEDWORDS Proline dehydrogenase； Chronic heart failure； Heart failure with reduced ejection fraction； Heart failure with mid-range ejection fraction； Heart failure with preserved ejection fraction

Objective To analyze the clinical data of one patient with inguinal seminoma and testicular seminoma，and to provide the references for the diagnosis and treatment of such patients. Methods The clinical data， imaging features， pathological manifestations， diagnosis， and treatment methods of one patient with primary inguinal seminoma were collected and the relevant literatures were reviewed. Results The patient， a 43-year-old male， was admitted to hospital due to the incidental discovery of a left inguinal mass. The specialized examination results showed a mass with relatively clear boundary， rough surface， palpable nodules， local fluctuation， limited mobility and no epidermal ulceration. The skin temperature was slightly elevated. The morphology and size of both testes were normal with no tenderness. The auxiliary examination results showed that alpha-fetoprotein （AFP） level was 2.3 mg·L^-1， beta-human chorionic gonadotropin （HCG） <0.1 IU·L^-1， and the lactate dehydrogenase （LDH） level was 254.31 IU·L^-1. The results of liver function， renal function， blood routine， and coagulation routine of the patient were all normal.The results of chest CT， ECG， cardiac ultrasound， liver-gallbladder-spleen-pancreas ultrasound， and retroperitoneal ultrasound showed no abnormalities. The preoperative biopsy pathology from the oncology department suggested seminoma. After comprehensive clinical analysis， it was decided to perform the inguinal mass excision and inguinal lymph node dissection. The postoperative pathology was consistent with the preoperative findings， confirming that seminoma with inguinal lymph node metastasis. The follow-up results at 1， 3， 6， and 8 months after operation showed the patient recovered well， and there were no discomfort and capable of light physical activities. The condition remained stable without progression， and the patient had undergone three sessions of radiochemotherapy in the Oncology Department. Conclusion Primary inguinal seminoma has a high cure rate， generally low malignancy， sensitivity to radiotherapy and chemotherapy， and relatively good prognosis.

Objective To discuss the clinical characteristics， intervention timing， diagnosis and treatment plan， and prognosis of one patient with immune thrombotic thrombocytopenic purpura （iTTP）， and to provide more clinical evidences for the precise diagnosis and treatment of this rare disease. Methods The clinical data of one patient with iTTP were collected， who had been previously misdiagnosed with acute infection after receiving a virus-inactivated vaccine. The data included clinical manifestations and ancillary examination informations. The relevant literatures were reviewed. Results The patient， a 60-year-old male， was admitted with “fever for 6 d.” The physical examination results showed scattered red maculopapules on the lower limbs， with only mild thrombocytopenia initially， and the admission PLASMIC score was 5 points. Initially diagnosed with acute infection， the patient was treated with anti-inflammatory， anti-infective， and corticosteroid therapies， but the response was poor. After one week， the re-evaluation results showed a significant decrease in the platelet count， progressing to severe thrombocytopenia， hematuria， dark-colored urine， and neurological and psychiatric symptoms as the disease progressed. The further examination results showed the PLASMIC score was increased to 7 points. After high suspicion of iTTP， therapeutic plasma exchange （TPE） was initiated immediately. The a disintegrin and metalloproteinase with thrombospondin type 1 motif member 13（ADATMS13） activity level was<1% during treatment， and the test for ADAMTS13 inhibitors was positive. The genetic testing results revealed a missense mutation in an iTTP-susceptible gene. After a confirmed diagnosis， the patient was treated regularly with intravenous rituximab， completing four treatment cycles， and followed up to the present； the treatment deemed effective. Conclusion iTTP is often delayed in the diagnosis due to atypical initial clinical symptoms. Once suspected， the treatment based on TPE and glucocorticoids should be initiated immediately. New drugs like rituximab provide a multidisciplinary treatment strategy option for iTTP.

Objective To discuss the association between occupational acid fog exposure and accelerated biological aging of the workers， and to clarify its related risk factors. Methods A total of 341 male workers exposed to occupational acid fog and 201 male workers without occupational exposure were selected as the study subjects, and they were divided into exposure group and control group, respectively. The general informations of the subjects in two groups were collected through questionnaires and physical examinations. The levels of red blood cell count（RBC）, platelet count（PLT）, albumin（ALB）, urea（Urea）, creatinine（CR）, triglycerides（TG）, total cholesterol（TC）, glycated hemoglobin（HBA1c）, and high-sensitivity C-reactive protein（Hs-CRP） in serum of the subjects in two groups were detected. The Klemera-Doubal method （KDM） was used to construct the composite aging measure, KDM-biological age（BA）（KDM-BA）. The model parameters were trained using samples from the 2009 China Health and Nutrition Survey（CHNS） Database to calculate the BA acceleration of the subjects in two groups； stratified analysis based on the population characteristics was conducted to analyze the BA of the subjects in two groups with different population characteristics； generalized linear model was used to analyze the factors influencing BA acceleration due to acid fog exposure. Results The model parameters were trained using samples from the 2009 CHNS Database, including 8 133 cases aged 20-79 years, of which 3 788 were male. The levels of Urea, CR, HBA1c, ALB, and TC, as well as systolic blood pressure (SBP), total working years, sleep duration, and body mass index（BMI） of the subjects between two groups had significant differences（P<0.05）. Compared with control group, the BA acceleration of the subjects in exposure group was significantly increased （P<0.05）. In entire population and exposure group, the BA acceleration in the smokers was significantly higher than that in the non-smokers （P<0.05）. In entire population, control group, and exposure group, the BA accelerations of the subjects in different BMI groups were significantly decreased with the increase of BMI （P<0.05）. Compared with control group, the BA acceleration of the subjects in exposure group was significantly increased（P<0.05）， including those under 40 years old, with total working years of 4-7 years, Han nationality, unmarried, smokers, and sleep duration 6-7 h, and with overweight. Acid fog exposure, smoking, and BMI were associated with the BA acceleration （β=0.72, 95% CI： 0.24—1.21； β=0.59, 95% CI： 0.11—1.06； β=-0.29, 95% CI： -0.35—-0.22）. Conclusion Occupational acid fog exposure may accelerate the biological aging in the workers, and acid fog is a risk factor to accelerate the biological aging of the body.

The gut microbiota is a vast microbial ecosystem， specifically present in the organism and plays an important regulatory role in the body’s health or disease state together with its metabolites. After spinal cord injury， the complex pathophysiology at the site of trauma makes axonal regeneration difficult， and the autonomic motor dysfunction induced by spinal cord injury disrupts gastrointestinal function and causes gut microbiota imbalance. The previous clinical outcomes of neurorepair strategies after spinal cord injury have not been ideal. The dysregulated gut microbiota and neuroinflammation after spinal cord injury are closely associated with the prognosis of the patients. The potential mechanisms by which the gut microbiota may influence the neuroinflammation after spinal cord injury may include the activation of gut-associated lymphoid tissue and disruption of the intestinal barrier by the imbalanced microbiota， and gut microbiota and its metabolites such as lipopolysaccharides（LPS）， short chain fatty acids（SCFAs）， 5-hydroxytryptamine（5-HT）， and tryptophan， as well as immune cells， inflammatory factors， and neurotransmitters the local inflammatory response in the spinal cord through the circulatory system. This paper revews the studies on the changes in gut microbiota after spinal cord injury and their effects on the spinal cord neuroinflammation， providing new targets and new ideas for improving the neuroinflammation after spinal cord injury.

Metabolic syndrome （MS） is a complex syndrome based on metabolic disorders in the human body， and is a risk factor for cardiovascular diseases and even certain tumors， with a complicated etiology and unclear pathogenesis. Helicobacter pylori （Hp） is one of the most common pathogenic bacteria， closely associated with the occurrence and development of various diseases. Currently， there are numerous studies both domestically and internationally on the relationship between Hp and MS and its components. Most studies suggest that there is an association between Hp and MS and Hp influences the occurrence and development of MS through multiple pathways. Eradicating Hp may become a new option for treating MS. Based on recent studies from both domestic and international sources， this paper discusses the association between Hp and MS， analyzes the effects of Hp on obesity， blood glucose， blood lipids， and blood pressure， and aims to provide new ideas for the prevention and treatment of MS.

Lipoprotein（a） ［Lp（a）］ is a highly polymorphic lipoprotein molecule composed of apolipoprotein A （apo A）， apo B100， and cholesterol ester core； calcific aortic valve stenosis （CAVS） is a multifactorial valvular heart disease influenced by both environmental and genetic factors. Lp（a） is an independent risk factor for CAVS and can increase the onset risk of CAVS. Lp（a） plays an important role in the treatment of CAVS. Although surgery is currently the main clinical treatment for CAVS， with further exploration into its pathological mechanism， drug therapy targeting Lp（a） has emerged as a new method. This paper reviews studies about the structure and characteristics of Lp（a）， its role in the occurrence and development of CAVS， treatment options related to hyperlipoprotein Aemia， and the studies preventing and treating CAVS by combating inflammation， and enhances the clinicians’ understanding and awareness of hyperlipoprotein Aemia and provides new insights for the prevention and targeted drug therapy of CAVS.

Dental pulp stem cells （DPSCs） are a type of mesenchymal stem cells with broad application potential. Due to their multipotent differentiation capabilities and ease of access， DPSCs have become a focus of research in the field of ophthalmology. In recent years， DPSCs have shown the potential clinical applications in the treatment of corneal epithelial injuries and retinal degenerative diseases. DPSCs can promote the corneal epithelial regeneration and reconstruction by differentiating into the corneal epithelial cells and inhibiting the M1 macrophages. Additionally， DPSCs can differentiate into the retinal photoreceptor-like cells and retinal ganglion-like cells， replace original retinal neurons， secrete neurotrophic factors to mediate injury repair， promote retinal regeneration， and improve the original function of the retina. This article systematically reviews the relevant domestic and international literatures in recent years， discussing the biological characteristics of DPSCs and the research progress in their applieation in the treatment of corneal and retinal diseases， with the aim of providing insights and strategies for the study of DPSCs in translational medicine and ophthalmic-related diseases.KEYWOEDS Dental pulp stem cells； Eye； Cornea； Retina； Paracrine

Tooth ankylosis is a clinical condition where the tooth cementum directly fuses with the surrounding alveolar bone， leading to functional and aesthetic defects. The etiology involves genetic， metabolic， and local stimulation factors. The diagnosis of tooth ankylosis requires a combination of clinical manifestations and imaging examinations to improve the diagnostic accuracy. The treatment of tooth ankylosis presents significant challenges. Orthodontic treatment combined with other disciplines offers a new， comprehensive treatment approach， integrating traditional orthodontic techniques with osteotomy， distraction osteogenesis， orthodontic bone traction， corticotomy， dislocation， and autologous tooth transplantation techniques. The treatment of tooth ankylosis requires the cooperation of multiple disciplines， and the experts from orthodontics， oral surgery， and oral medicine collaborate to develop the optimal treatment plan. This comprehensive treatment method achieves better outcomes compared with traditional treatments. This review discusses the etiology， diagnosis， and orthodontic combined multidisciplinary treatment methods of tooth ankylosis， analyzes the advantages and disadvantages of various treatment options， evaluates the efficacy and risks， and provides new perspectives for the treatment of tooth ankylosis.