Objective To discuss the effect of nobiletin （NOB） on airway hypersecretion in the rats with chronic obstructive pulmonary disease （COPD）， and to clarify the molecular mechanism of endoplasmic reticulum stress （ERS） signaling pathway regulation. Methods A total of 45 SD rats were randomly divided into control group， model group， NOB group， endoplasmic reticulum stress inhibitor 4-phenylbutyric acid （4-PBA） group， and NOB+4-PBA group， and there were 9 rats in each group. Except for control group， all the rats in the other groups were exposed to smoke and intratracheally injected with lipopolysaccharide （LPS） to establish the COPD models. After successful modeling， the rats were injected with the corresponding drugs （50 mg·kg^-1·d^-1 NOB or 0.35 g·kg^-1·d^-1 4-PBA） once daily for 4 consecutive weeks. After the drug intervention， the pulmonary function indexes ［dynamic compliance （Cdyn） and airway resistance （RL）］ and arterial blood gase indexes ［partial pressure of carbon dioxide （PaCO₂） and oxygen partial pressure （PaO₂）］ of the rats in various groups were detected by small animal ventilator and blood gas analyzer. Hematoxylin and eosin （HE） staining was used to observe the hyperplasia of the goblet cells in lung tissue of the rats in various groups； enzyme-linked immunosorbent assay （ELISA） method was used to detect the levels of mucin 5AC （MUC5AC）， interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， and interleukin-6 （IL-6） in bronchoalveolar lavage fluid （BALF） of the rats in various groups；real-time fluroscence quantitative PCR （RT-qPCR） method was used to detect the expression level of MUC5AC mRNA in lung tissue of the rats in various groups；Western blotting method was used to detcet the protein expression levels of MUC5AC and ERS-related proteins inositol-requiring enzyme 1α （IRE1α）， activating transcription factor 6 （ATF6）， CCAAT-enhancer-binding protein homologous protein （CHOP）， and glucose-regulated protein 78 （GRP78） in lung tissue of the rats in various groups. Results Compared with control group，the number of airway goblet cells of the rats in model group was increased（P<0.05）， the Cdyn and PaO₂ levels were decreased （P<0.05）， and the levels of RL and PaCO₂ were increased （P<0.05）； the levels of MUC5AC， IL-1β， TNF-α， and IL-6 in BALF and the expression levels of MUC5AC， IRE1α， ATF6， CHOP， and GRP78 proteins in lung tissue were increased （P<0.05）. Compared with model group， the lung function， arterial blood gas indexes， and proliferation of airway goblet cells of the rats in NOB group， 4-PBA group， and NOB+4-PBA group were significant improved， the levels of IL-1β， TNF-α， and IL-6 in BALF and the expression levels of MUC5AC， IRE1α， ATF6， CHOP， and GRP78 proteins in lung tissue were increased（P<0.05）； the rats in NOB+4-PBA group exhibited the best alleviation effect. Conclusion NOB can effectively alleviate the airway mucus hypersecretion in the rats with COPD， and its mechanism may be related to the suppression of MUC5AC overexpression mediated by ERS.

Objective To prepare the anti-herpes simplex virus-1 （HSV-1） egg yolk antibody （IgY） and discuss its biological activity， and to clarify its anti-HSV-1 capability. Methods The inactivated HSV-1 vaccine was prepared， the high egg-producing hens were immunized by multi-site injection method in the chicken breast， and IgY was purified by polyethylene glycol-6000 （PEG-6000） method；the titer， purity， and protein levels of IgY were detected by indirect ELISA method， sodium dodecyl sulfate-polyacrylamide gel electrophoresis （SDS-PAGE） technology， and protein concentration quantification kits； the ability of IgY to neutralize the virus was detected by indirect ELISA method； the virus blocking rate was detected.The Vero cells were used as the target cells for viral infection. The cytopathic effect （CPE） at different antibody concentrations （0.015 60， 0.031 25， 0.062 50， 0.125 00， 0.250 00， 0.500 00， and 1.000 00 g·L?1） was detected；the in vitro anti-HSV-1 capacity of IgY was detected according to the degrees of CPE. Results The titer of the prepared IgY was gradually increased with the extension of immunization time. The titer of the IgY stabilized at 1/1 024 000； the clear bands for the IgY light and heavy chains were observed； the average protein level of IgY was 11.544 g·L?1； the blocking rate of IgY against HSV-1 could reach up to 77.90%； the degree of HSV-1 induced CPE was decreased with the increasing of the antibody concentrations， and when the concentration of IgY reached 0.500 00 g·L?1 or higher， less than 25% of the cells （even no cells） exhibited cytopathic changes. Conclusion Anti-HSV-1 IgY is successfully prepared， and this antibody has significant neutralizing and blocking capabilities and in vitro inhibitory activity against HSV-1， which can be utilized for the development of drugs and diagnostic methods.

Objective To discuss the effects of microRNA-126 （miR-126） over-expression and disintegrin and metalloproteinase 9 （ADAM9） gene silencing on the biological behavior of the gastric cancer cells， and to clarify the mechanism. Methods The human poorly differentiated gastric adenocarcinoma SGC-7901 cells and human normal gastric mucosa epithelial NGEC cells were cultured in vitro. The total RNA was extracted from the cells， and the expression levels of miR-126 and ADAM9 mRNA in both types of cells were detected by real-time fluorescence quantitative PCR （RT-qPCR）method. The SGC-7901 cells at logarithmic growth phase were divided into miR-126 over-expression group （miR-126-OE group） and ADAM9 gene silencing group （ADAM9 siRNA group）. The transfection with miR-126 mimics （miR-126） mimics and ADAM9 RNA oligonucleotides were conducted by Lipofectamine^TM 2000， and the corresponding negative control group was established； MTT assay was used to detect the proliferation activities of the cells in various groups； cell wound assay was used to detect the migration rate of the cells in various groups；Transwell chamber assay was used to detect the the numbers of migration and invasion cells in various groups； Western blotting method was used to detect the expression levels of E-cadherin， N-cadherin， and vimentin proteins in the cells in various gorups. The miR-126 target genes were predicted by TargetScan website， and the targeting regulatory relationship between miR-126 and ADAM9 was confirmed by dual-luciferase reporter assay. The expression levels of ADAM9 mRNA and protein in the SGC-7901 cells after transfected with miR-126 mimics were detected by RT-qPCR and Western blotting methods. Results The RT-qPCR results showed that compared with human normal gastric mucosa epithelial NGEC cells， the expression level of miR-126 in the gastric cancer SGC-7901 cells was significantly decreased （P<0.05）， while the expression level of ADAM9 mRNA was significantly increased （P<0.05）. The MTT assay results showed that after 48 and 72 h of over-expressing miR-126 or silencing the ADAM9 gene in the SGC-7901 cells， compared with the corresponding negative control group， the proliferation activities of the cells in both miR-126-OE and ADAM9 siRNA groups were significantly decreased （P<0.05 or P<0.01）. The cell wound assay results indicated that compared with the corresponding negative control group， the migration rates of the cells in both miR-126 OE and ADAM9 siRNA groups 48 h after transfection were significantly decreased （P<0.05）. The Transwell chamber assay results showed that the numbers of migration and invasion cells in both miR-126-OE and ADAM9 siRNA groups were significantly lower than those in corresponding negative control group （P<0.05 or P<0.01）.The Western blotting method results showed that compared with the corresponding negative control groups， the expression level of E-cadherin protein in the cells in miR-126-OE and ADAM9 siRNA groups were significantly increased （P<0.05 or P<0.01）， while the expression levels of N-cadherin and vimentin proteins were significantly decreased （P<0.05 or P<0.01）. The target prediction results showed that the 3'-UTR of ADAM9 contains nucleotide sequences complementary to miR-126-3p. The dual-luciferase reporter assay results showed that ADAM9 was a downstream target gene negatively regulated by miR-126. Compared with mimics NC group， the expression levels of ADAM9 mRNA and protein in the SGC-7901 cells after transfected with miR-126 mimics for 48 h were decreased （P<0.05 or P<0.01）. Conclusion The gastric cancer SGC-7901 cells are characterized by low expression of miR-126 and high expression of ADAM9 gene. Over-expression of miR-126 can inhibit the proliferative activity， migration， and invasion capabilities of the gastric cancer SGC-7901 cells； the mechanism may be related to the negative regulation of ADAM9 by miR-126 and the inhibition of epithelial-mesenchymal transition （EMT） process in the gastric cancer cells.

Objective To observe the improvement effect of total flavonoids of Sophora flavescens （SF） on the oxidative stress in the Leydig’s cells TM3 of the mice induced by lead acetate （LA） and their influences on the Keap1/Nuclear factor E2-related factor 2 （Nrf2） signaling pathway， and to discuss the potential mechanism of the improvement effect of SF on oxidative stress in the LA-induced TM3 cells. Methods The survival rates of the TM3 cells after treated with various concentrations of LA （0， 10， 20， 50， and 80 μmol·L^－1） and SF （0， 12.5， 25.0， 50.0， and 80.0 mg·L^－1） for 24 h were detected by MTT assay. The TM3 cells were randomly divided into blank control group， LA group， LA + low dose of SF group， LA+middle dose of SF group， and LA+high dose of SF group. Except for blank control group， the TM3 cells in the other groups were induced with 50 μmol·L^－1 LA for 24 h to establish the oxidative stress model. The TM3 cells in LA+low dose of SF group， LA+middle dose of SF group， and LA+ high dose of SF group were then treated with 12.5， 25.0， and 50.0 mg·L^－1 SF， respectively. The survival rate of the TM3 cells was detected by MTT assay； the activities of superoxide dismutase （SOD） and levels of malondialdehyde （MDA） in the TM3 cells in various groups were detected by WST-1 assay and thiobarbituric acid （TBA） method； the expression levels of Nrf2， Keap1， and Caspase-9 proteins in the TM3 cells in various groups were detected by detected by Western blotting method. Results The MTT assay resuts showed that compared with 0 μmol·L^－1 LA group，the survival rates of the TM3 cells in 10， 20， 50， and 80 μmol·L^－1 LA groups were decreased （P<0.01）； compared with 0 mg·L^－1 SF group，the survival rates of the TM3 cells in 12.5， 25.0， 50.0， and 80.0 mg·L^－1 SF groups were significantly decreased （P<0.05 or P<0.01）； compared with LA group， the survival rates of the TM3 cells in LA+low dose of SF group， LA+middle dose of SF group， and LA+high dose of SF group were significantly increased （P<0.01）. The WST-1 assay and TBA method results showed that compared with LA group，the SOD activities and MDA levels in the TM3 cells in LA+low dose of SF group， LA+middle dose of SF group， and LA+high dose of SF group were significantly decreased （P<0.01）.The Western blotting results showed that compared with LA group， the expression levels of Nrf2 protein in the TM3 cells in LA+low dose of SF group， LA+middle dose of SF group， and LA+high dose of SF group were increased（P<0.01），and the expression levels of Keap1 and Caspase-9 proteins were decreased （P<0.01）. Conclusion SF has certain improvement effect on the oxidative stress in the LA-induced TM3 cells and can reduce the expression level of Keap1 protein and increase the expression level of Nrf2 protein in the TM3 cells.

Objective To discuss the effect of rubusoside （RUB） on the spinal cord injury （SCI） and neuroinflammation in the mice， and to clarify its mechanism. Methods A total of 48 female Kunming mice were randomly divided into sham operation group， SCI group， SCI+low dose of RUB group， and SCI+high dose of RUB group， and there were 12 mice in each group. Spinal cord injury behavior （BBB） scoring method was used to assess the motor function of the hind limbs of the SCI mice； water content method was used to detect the spinal cord edema of the SCI mice； real-time fluorscence quantitative PCR （RT-qPCR） method was used to detect the expression levels of pro-inflammatory cytokines cyclooxygenase-2 （COX-2）， interleukin-1β （IL-1β）， and tumor necrosis factor-α （TNF-α） mRNA of the mice in various groups； ELISA method was used to detect the levels of inflammatory factors in serum of the mice in various groups； HE staining was used to observe the pathomorphology of spinal cord tissue of the mice in various groups；immunofluorescence was used to detect the activation of microglia in spinal cord tissue of the mice in various groups； Western blotting method was used to detect the expression levels of related proteins in spinal cord tissue of the mice in various groups. Results The BBB score analysis results showed that compared with sham operation group， the score of the mice in SCI group was decreased to 0； compared with SCI group， the BBB scores of the mice in SCI+low dose of RUB group and SCI+high dose of RUB group were gradually increased. The spinal cord tissue water content detection results showed that compared with sham operation group， the water content in spinal cord tissue of the mice in SCI group was significantly increased （P<0.01）； compared with SCI group， the water contents in spinal cord tissue of the mice in SCI+ low dose of RUB group and SCI+ high dose of RUB group were significantly decreased （P<0.01）. The RT-qPCR results showed that compared with sham operation group， the expression levels of COX-2， IL-1β， and TNF-α mRNA in spinal cord tissue of the mice in SCI group were significantly increased （P<0.001）. Compared with SCI group， the expression levels of COX-2，IL-1，and TNF-α mRNA in spinal cord tissue of the mice in SCI+ low dose of RUB group and SCI+ high dose of RUB group were significantly decreased （P<0.001）. The ELISA results showed that compared with sham operation group，the levels of IL-1β （P<0.01） and TNF-α （P<0.001） in serum of the mice in SCI group were increased； compared with SCI group， the levels of IL-1β and TNF-α in spinal cord tissue of the mice in SCI+ low dose of RUB group and SCI+ high dose of RUB group were decreased （P<0.001）.The Western blotting results showed that compared with sham operation group， the expression levels of nuclear factor kappa B （NF-κB） inhibitor protein-α （IκB-α）， phosphorylated IκB-α （p-IκB-α）， phosphorylated p65 （p-p65）， p65， phosphorylated p38 （p-p38）， phosphorylated extracellular regulated protein kinase （p-ERK）， and phosphorylated c-Jun N-terminal kinase （p-JNK） proteins in spinal cord tissue of the mice in SCI group were significantly increased （P<0.05 or P<0.001）； compared with SCI group， the expression levels of IκB-α， p-IκB-α， p-p65， p65， p-p38， p-ERK， and p-JNK proteins in spinal cord tissue of the mice in SCI+ low dose of RUB group and SCI+ high dose of RUB group were significantly decreased （P<0.001）. The HE staining results showed that compared with sham operation group， the spinal cord tissue of the mice in SCI group had loose organization with the formation of vacuoles， and a large necrotic cavity was present in the center of the spinal cord； compared with SCI group， the central necrotic cavity area of the mice in SCI+ low dose of RUB group and SCI+ high dose of RUB group was significantly decreased （P<0.05）.The immunofluorescence results showed that compared with sham operation group， the number of positive microglia cells in spinal cord tissue of the mice in SCI group was significantly increased （P<0.001）； compared with SCI group， the numbers of positive microglia cells in spinal cord tissue of the mice in SCI+ low dose of RUB group and SCI+ high dose of RUB group were significantly decreased （P<0.001）. Conclusion Rubusoside can improve the motor function impairment in the SCI mice，mitigate the neuroinflammation in spinal cord tissue，and inhibit the activation of microglia. The mechanism may be related to the downregulation of expression of proteins associated with the NF-κB and mitogen-activated protein kinase （MAPK） signaling pathways in spinal cord tissue.

Objective To construct the antibacterial hydrogel containing mesenchymal stem cell exosomes （MSC-Exo） and silver nanoparticles （AgNPs）， and to preliminarily clarify its antibacterial activity and proliferative effect on the human epidermal HaCaT cells， and to clarify its application potential in treatment of skin wound. Methods The basic hydrogel scaffold was fabricated using oxidized dextran （ODex） and modified hyaluronic acid （HA-ADH）. The AgNPs were synthesized with cetyltrimethylammonium bromide （CTAB） as the stabilizer， while the MSC-Exo was isolated from the cultured mesenchymal stem cells （MSC）. The AgNP/HA-ADH/ODex hydrogel and MSC-Exo@AgNP/HA-ADH/ODex hydrogel were constructed. The loading rate of the Msc-Exo was evaluated through BCA assay； dynamic light scattering （DLS）， Zeta potential measurement， and ultraviolet-visible spectroscope were used to analyze the morphology， the charged status of the AgNPs； fourier transform infrared spectra was used to detect the compositions of the HA-ADH/ODex hydrogel framework； transmission electron microscope （TEM）， nanoparticle tracking analysis， and Western blotting methods were used to identify the presence of MSC-Exo；the cell survival rates of the HA-ADH/ODex hydrogel and the AgNP/HA-ADH/ODex hydrogel， and the effect of MSC-Exo@AgNP/HA-ADH/ODex hydrogel on the proliferation rate of HaCaT cells were detected by CCK-8 assay；the antibacterial efficacy of the AgNP/HA-ADH/ODex hydrogel was detected by spread plate method and turbidity measurement. Results The DLS， Zeta potential， and ultraviolet-visible spectrum results showed the successful synthesis of uniform and positively charged AgNPs. The fourier transform infrared spectrometry results showed the HA-ADH/ODex hydrogel was properly formulated. The identification of MSC-Exo was verified by TEM， nanoparticle tracking， and Western blotting methods. The BCA assay results showed that the loading rate of the MSC-Exo on the MSC-Exo@AgNP/HA-ADH/ODex hydrogel was 77.8%.The CCK-8 assay results showed that the survival rates of the HaCaT cells were close to 100% when treated with the dilution ratios of 0，1， 2， 4， 8， 16， 32， and 64 of culture medium-extract of HA-ADH/ODex hydrogel. For the AgNP/HA-ADH/ODex hydrogel extracted with dilution ratio of 8 and above， the survival rates were greater than 85%. The spread plate method results showed that the antibe terial rates of AgNP/HA-ADH/ODex and MSC-Exo@AgNP/HA-ADH/ODex hydrogels against Staphylococcus aureus （S. aureus） and Escherichia coli （E. coli）， and the antibacterial rates against S.aureus and E. coli were （96.44±0.16）%， （96.62±0.16）% and （95.73±0.28）%， （95.58±0.14）%， respectively. The CCK-8 assay results showed that the MSC-Exo@AgNP/HA-ADH/ODex hydrogel promoted the proliferation of the HaCaT cells，and the proliferation rate was （115.00 ± 7.42）%. Conclusion The MSC-Exo@AgNP/HA-ADH/ODex hydrogel exhibits the excellent antibacterial properties and good ability to promote the proliferation of human epidermal HaCaT cells； it is a safe and effective bio-dressing for the treatment of skin wounds.

Objective To discuss the effect of gene silencing of targeting protein for Xenopus kinesin like protein 2 （TPX2） on the chemosensitivity of the resistant bladder cancer cell line T24/cisplatin（DDP）， and to clarify the mechanism. Methods The DDP-resistant cell line T24/DDP was established by DDP concentration gradient induction method， and the cells were divided into T24 cell group and T24/DDP cell group. MTT method was used to detect the proliferation activities of the cells in various groups； the resistance index （RI） was calculated based on the half maximal inhibitory concentration （IC₅₀） value；real-time fluorescence quantitative PCR （RT-qPCR） and Western blotting methods were used to detect the expression levels of TPX2 mRNA and protein in the cells in various groups； small interfering RNA （siRNA） was used to silence the TPX2 gene expression in the T24/DDP cells.The cells were divided into blank control group， negative control siRNA （si-NC） group， TPX2 silenced （si-TPX2） group， si-NC+DDP （2 mg·L^-1 DDP） group，and si-TPX2+DDP （2 mg·L^-1 DDP） group. RT-qPCR and Western blotting methods were used to detect the expression levels of TPX2 mRNA and protein in the cells in various groups； MTT method was used to detect the proliferation activity of the cells in various groups； flow cytometry was used to detect the apoptotic rates of the cells and percentages of the cells at G₂/M phase in various groups； Transwell chamber assay was used to detect the numbers of migration and invasion cells； Western blotting method was used to detect the expression levels of Wnt/β-catenin signaling pathway-related proteins such as β-catenin， P-glycoprotein （P-gp）， zinc finger protein transcription factor 1 （Snail1）， and Survivin proteins in the cells in various groups. Results The resistant bladder cancer cell line T24/DDP was successfully established with the RI value of 8.76. Compared with T24 cell group， the expression levels of TPX2 mRNA and protein in the T24/DDP cells were significantly increased （P<0.01）. Compared with blank control group and si-NC group， the expression levels of TPX2 mRNA and protein in the T24/DDP cells in si-TPX2 group were significantly decreased （P<0.01）， and the IC₅₀ value of DDP was significantly decreased （P<0.01）. Compared with si-NC group， the apoptotic rate of the cells and the percentage of the cells at G₂/M phase in si-TPX2 group was significantly increased （P<0.01）， and the numbers of migration and invasion cells were significantly decreased （P<0.01）， and the expression levels of β-catenin， P-gp， Snail1， and Survivin proteins in the T254/DDP cells were also significantly decreased （P<0.01）. Compared with si-NC+DDP group， the apoptotic rate of the cells and percentage of the cells at G₂/M phase in si-TPX2+DDP group were significantly increased （P<0.01）， and the numbers of migration and invasion cells were significantly decreased （P<0.01）， and the expression levels of β-catenin， P-gp， Snail1， and Survivin proteins in the T24/DDP cells were significantly decreased (P<0.01). Conclusion Gene silencing of TPX2 enhances the chemosensitivity of the resistant bladder cancer cell line T24/DDP to DDP by inhibiting the Wnt/β-catenin signaling pathway.

Objective To discuss the effects of genomic instability and MYC gene mutation of non-small cell lung cancer （NSCLC） A549 cells on the resistance of gemcitabine， and to clarify the mechanism. Methods The A549 cells were continuously treated with 2 mg·L^－1 of gemcitabine （A549 group） to establish the resistant cell line A549R （A549R group）， and si-NC and si-MYC were transfected into the A549R cells to regarded as si-NC A549R group and si-MYC A549R group， respectively. CCK-8 assay was used to detect the inhibitory rates of the cells in various groups after treated with various concentrations of gemcitabine （0， 1， 2， 4， 8， 16， and 32 mg·L^－1）；flow cytometry was used to detect the apoptotic rates of the cells in various groups； transcriptome sequencing technology （RNA-seq） and Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathway enrichment analysis were used to identify the differentially expressed genes in the A549 and A549R cells； real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of muts homology 2 （MSH2）， muts homology 6 （MSH6）， and recombinant DNA repair protein RAD50（RAD50） in the A549 and A549R cells； Western blotting method was used to detect the expression levels of genome instability-related proteins MYC proto-oncogene （MYC） and phosphorylated H2AX （γH2AX） in the cells in various groups； chromatin immunoprecipitation （ChIP） was used to detect the enrichment of RNA pol Ⅱ and γH2AX on the MYC gene； PCR method was used to amplify and detect the mutations in MYC gene in the A549R cells. Results Compared with A549 group， the inhibitory rates of the A549R cells in A549R group treated with 2， 4， 8， 16， and 32 mg·L^－1 gemcitabine were decreased， and the apoptotic rate of the cells after treated with 8 mg·L^－1 gemcitabine was decreased （P<0.05）. Compared with A549 group， a total of 234 mRNAs in the cells in A549R group were upregulated and 205 mRNAs in the cells were downregulated， and the expression levels of mismatch repair-related genes （MSH2 and MSH6）， RAD50， and MYC were significantly increased （P<0.05）. The KEGG signaling pathway enrichment analysis showed that the upregulated genes were mainly involved in non-homologous end-joining， mRNA surveillance pathway， and DNA replication signaling pathways. Compared with A549 group， the expression levels of MYC and γH2AX proteins in the cells in A549R group were increased （P<0.05）.The ChIP assay results showed that the enrichment of RNA pol Ⅱ and γH2AX at the MYC transcription start site and exon 2， with a G254A mutation identified in MYC exon 2 was increased. Compared with si-NC A549R group， the expression levels of MYC mRNA and protein in the cells in si-MYC A549R group were decreased （P<0.05）， the inhibitory rates of the cells after treated with 2， 4， 8， 16， and 32 mg·L^－1 gemcitabine were increased（P<0.05）， and the apoptotic rates were increased （P<0.05）. Conclusion The genome instability of the A549R NSCLC cells resistant to gemcitabine is increased， and the mutations and amplification occur in the MYC gene. Knockdown of MYC can restore the sensitivity of A549R cells to gemcitabine.

Objective To discuss the effect of sodium tanshinone ⅡA sulfonate （STS） on the function of human umbilical vein endothelial cells （hUVECs） after treated with uremic toxin， and to clarify its mechanism. Methods The hUVECs were passaged and divided into blank control group， uremic toxin-stimulation group， uremic toxin + STS group， and uremic toxin + STS + extracellular signal-regulated kinase （ERK） inhibitor group. The concentration of STS used in the last two groups was 10 mg·L^-1. The shear stress stimulation at 12 dyn·cm^-2 was applied to the cells in various groups. The proliferation activities of the cells in various groups were detected by CCK-8 assay； the expression levels of ERK， nuclear factor kappa B （NF-κB）， and type Ⅰ collagen proteins in the cells in various groups were detected by Western blotting method；the expression levels of ERK， NF-κB， and type Ⅰ collagen mRNA in the cells in various groups were detected by real-time fluorescence quantitative PCR （RT-qPCR） method；the apoptotic rates the cells in various groups were detected by TUNEL method. Results The CCK-8 assay results showed that after treated with shear stress， the probiferation activitres of the cells in uremic toxin-stimulation group and uremic toxin + STS + ERK inhibitor group were lower than that in uremic toxin + STS group （P<0.01）. The Western blotting results showed that compared with uremic toxin group， the expression levels of ERK， NF-κB， and type Ⅰ collagen proteins in the cells in uremic toxin + STS group were increased （P<0.01）. After inhibiting the ERK pathway， compared with blank control group， uremic toxin group， and uremic toxin + STS group， the expression levels of ERK， NF-κB， and type Ⅰ collagen proteins in the cells in uremic toxin + STS + ERK inhibitor group were significantly decreased （P<0.01）. The RT-qPCR results showed that compared with uremic toxin group， the expression levels of ERK， NF-κB， and type Ⅰ collagen mRNA in the cells in uremic toxin + STS group were increased （P<0.01）. After inhibiting the ERK signaling pathway， compared with blank control group， uremic toxin group， and uremic toxin + STS group， the expression levels of ERK， NF-κB， and type Ⅰ collagen mRNA in the cells in uremic toxin + STS + ERK inhibitor group were significantly decreased （P<0.01）.The TUNEL method detection results showed that the apoptotic rate in the cells in uremic toxin + STS group was lower than those in uremic toxin-stimulation group and uremic toxin + STS + ERK inhibitor group （P<0.05）. Conclusion A certain concentration of STS can improve the proliferation of the endothelial cells and reduce the apoptosis of the cells after treated with uremic toxins by modulating the expressions of NF-κB and type Ⅰ collagen mRNA and proteins through the ERK signaling pathway.

Objective To discuss the effect of silencing Forkhead box K1 （FOXK1） on the proliferation， migration， and invasion of the gastric cancer HGC-27 cells， and to clarify the possible mechanism. Methods The expression levels of FOXK1 mRNA in gastric cancer tissue and normal gastric tissue were consulted based on the Gene Expression Profiling Interactive Analysis （GEPIA） Database； the synthetic si-FOXK1 was used to transfect the gastric cancer HGC-27 cells in vitro， and the cells were divided into blank control group， nc-FOXK1 group， and si-FOXK1 group. Western blotting method was used to detect the transfection efficiencies of the cells in various groups； MTT assay was used to detect the proliferation abilities of the HGC-27 gastric cancer cells in various groups after transfected with si-FOXK1. The clone formation assay was used to detect the number of clone-forming of the HGC-27 gastric cancer cells after transfected with si-FOXK1；wound healing assay was used to detect the migration rates of the gastric cancer HGC-27 cells in various groups after transfected with si-FOXK1；Transwell chamber assay was used to detect the numbers of migration and invasion cells in various groups；Western blotting method was used to detect the expression levels of NF-κB pathway-related proteins ［nuclear factor κB p65 （NF-κB p65） and phosphorylated NF-κB p65 （p-NF-κB p65）］ in the gastric cancer HGC-27 cells in various groups after transfected with si-FOXK1. Results The GEPIA Database results showed that compared with normal gastric tissue， the expression level of FOXK1 mRNA in gastric cancer tissue was increased （P<0.05）. The Western blotting method results showed that the expression level of FOXK1 protein in the gastric cancer HGC-27 cells was higher than that in normal gastric mucosal GES-1 cells （P<0.01）. Compared with blank control group and nc-FOXK1 group， the expression level of FOXK1 protein in the cells in si-FOXK1 group was significantly decreased （P<0.01）. The MTT assay results showed that compared with blank control group and nc-FOXK1 group，the proliferation ability of the gastric cancer HGC-27 cells in si-FOXK1 group was decreased （P<0.05）. The clone formation assay results showed that compared with blank control group and nc-FOXK1 group， the number of clone-forming of the gastric cancer HGC-27 cells in si-FOXK1 group was decreased （P<0.05）. The cell scratch healing results showed that compared with blank control group and nc-FOXK1 group， the migration rate of the gastric cancer HGC-27 cells in si-FOXK1 group was decreased （P<0.05）.The Transwell chamber assay results showed that compared with blank control group and nc-FOXK1 group， the number of invasion cells of the gastric cancer HGC-27 cells in si-FOXK1 group was decreased （P<0.05）. The Western blotting results showed that compared with blank control group and nc-FOXK1 group， the expression level of p-NF-κB p65 protein in the cells in si-FOXK1 group was decreased （P<0.05）. Conclusion FOXK1 is highly expressed in the gastric cancer HGC-27 cells， and silencing FOXK1 can inhibit the proliferation， migration， and invasion abilities of the HGC-27 gastric cancer cells； its mechanism is possibly associated with the NF-κB pathway.

Objective To analyze the expression， clinical significance， and relationship with tumor immune infiltration of serine/arginine-rich splicing factor 9 （SRSF9） in head and neck squamous cell carcinoma （HNSCC） by bioinformatics methods， and to discuss its mechanism. Methods The expression of SRSF9 in HNSCC and its relationship with clinical pathologic characteristics and prognosis of the patients were analyzed by The Cancer Genome Atlas （TCGA） Database， GSE30784 and GSE13601 datasets in Gene Expression Omnibus （GEO） Database， Clinical Proteomic Tumor Analysis Consortium （CPTAC） Database， Gene Expression Profiling Interactive Analysis （GEPIA） Database， and Kaplan-Meier plotter Database；the variations of SRSF9 gene were examined through cBioPortal Database； ESTIMATE algorithm and The Tumor Immune Estimation Resource （TIMER） Database were used to assess the correlation between SRSF9 expression and tumor microenvironment， as well as tumor immune cell infiltration； LinkedOmics Database was used to analyze the SRSF9 co-expressed genes and their regulatory pathways in the HNSCC；the TCGA SpliceSeq Database was used to analyze the variable splicing events regulated by SRSF9 in the HNSCC；Gene Ontology （GO） functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathway enrichment analysis were conducted on the target genes. Results The analysis results from TCGA Database and GSE30784 and GSE13601 datasets in GEO Database showed that the expression level of SRSF9 mRNA in the HNSCC tissue was significantly higher than that in adjacent normal tissue （P<0.01）.The CPTAC Database analysis results showed that compared with normal tissue， the expression level of SRSF9 protein in the HNSCC tissue was significantly increased （P<0.001）.The SRSF9 mRNA expression was associated with pathological grading （P=0.004） and HPV infection （P=0.031） in the patients with HNSCC according to TCGA Database analysis. The GEPIA and Kaplan-Meier plotter Database analysis results showed that high expression of SRSF9 mRNA in the HNSCC tissue was correlated with poorer overall survival （OS） of the patients ［hazard ratio （HR） = 1.40， P=0.019； HR=1.55， P=0.003］. The cBioPortal Database analysis results showed that copy number variation （CNV） of SRSF9 gene occurred in 26.85% in HNSCC， and CNV was positively correlated with SRSF9 mRNA expression levels （r=0.44， P<0.001）. The ESTIMATE algorithm analysis results showed that high expression of SRSF9 mRNA group had lower stromal and immune score， and higher tumor purity than those in low expression of SRSF9 mRNA group（P<0.001）. The TIMER Database analysis results showed there was a positive correlation between the expression of SRSF9 mRNA and CD4+ T lymphocyte infiltration （r=0.186， P<0.001）， and there were negative correlations between the expression level SRSF9 mRNA and B lymphocyte， CD8+ T lymphocyte， and dendritic cell infiltrations （r=－0.269， P<0.001； r=－0.353， P<0.001； r=－0.304， P<0.001）. The co-expressed gene enrichment analysis results showed upregulation of genes related to ribosomes， spliceosomes， and metabolic pathways， and downregulation of genes associated with focal adhesions， cytokines， and cell adhesion molecules. The main pathways involved in SRSF9-related variable splicing target genes were lipid metabolism， glucagon， and tight junctions. Conclusion SRSF9 is highly expressed in HNSCC and is associated with poor prognosis and tumor microenvironment immune cell infiltration， suggesting its potential as a molecular target for the diagnosis， prognosis assessment， and treatment of HNSCC.

Objective To identify the key microRNAs （miRNAs） and their target genes associated with the prognosis of triple-negative breast cancer （TNBC） and to discuss their roles in regulatory biological functions and signaling pathways， and to provide the theoretical basis for the selection of prognostic biomarkers for the patients with TNBC. Methods The differentially expressed miRNAs between the TNBC patients and non-TNBC patients were selected based on The Cancer Genome Atlas （TCGA） Database； survival analysis was used to clarify the miRNAs related to the prognosis of the patients；miRDB and miRWalk3.0 online Databases were used to screen for the miRNA target gene； Cytoscape 3.8.2 software was used to elucidate the miRNA-messenger RNA （mRNA） regulatory network；the limma package in R software was used to screen the differentially expressed miRNAs of the TNBC patients and non-TNBC patients； the clusterProfiler package in R software was used to analyze the biological functions and pathways involved by the target genes. Results The survival analysis results showed that miR-9， miR-17， miR-31， miR-146a， miR-188， and miR-190b were associated with the prognosis of the TNBC patients， and low expression levels of these miRNAs were correlated with the poorer prognosis of the patients. A total of 224 target genes regulated by these six miRNAs were identified. The Gene Ontology （GO） functional enrichment analysis results showed that the target genes were primarily involved in the development of mammary alveoli， positive regulation of DNA transcription， and angiogenesis.The Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis results showed that the target genes were mainly enriched in cytokine-cytokine receptor interaction and insulin secretion signaling pathways. The network analysis results identified the key genes in the regulatory network， including solute carrier family 24 member 2 （SLC24A2）， vav guanine nucleotide exchange factor 3 （VAV3）， tripartite motif containing 36 （TRIM36）， synaptotagmin 1 （SYT1）， pleckstrin and Sec7 domain containing 3 （PSD3）， peroxisome proliferator-activated receptor α （PPARA）， RNA polymerase Ⅲ subunit G （POLR3G）， pleomorphic adenoma gene 1 （PLAG1）， ubiquitin associated and SH3 domain containing B （UBASH3B）， and SH3 domain and tetramerization domain containing 2 （SH3TC2）； the interactions of miRNA-mRNA of miR-9-SYT1， miR-9-KIF13B， miR-9-KITLG， miR-17-SLC24A2， miR-31-SLC24A2，miR-146a-SYT1， miR-146a-KIF13B， miR-188-SLC24A2， and miR-188-SLC24A2 were the closest. Conclusion MiR-9， miR-17， miR-31， miR-146a， miR-188， miR-190b and their target genes are involved in physiological processes such as mammary alveolar development and angiogenesis， which are closely associated with the poor prognosis of the patients with TNBC.

Objective To discuss the expression of ubiquitinated histone （H2AX） in the prostate cancer （PCa） and adjacent benign prostate tissues， and its relationship with the clinicopathological parameters of the PCa patients，and to clarify the effect of the novel histone deacetylase inhibitor （HDACi） CUDC-101 on the DNA damage， migration， and epithelial-mesenchymal transition （EMT） in PCa. Methods The expression levels of H2AX mRNA in various cancer tissues were retrieved from The Cancer Genome Atlas （TCGA） and UALCAN Databases to analyze the expression differences between PCa and normal prostate tissues and its connection with the clinical prognosis of the patients with PCa； immunohistochemistry method was used to detect the expression of H2AX protein in PCa tissue and adjacent benign prostate tissue， and its relationship with the clinicopathological parameters of the PCa patients was analyzed. The DU145 cells were cultured in vitro and divided into control group， 5% FBS group， 5% FBS+100 μmol·L^-1 CUDC-101 group， and 5% FBS+200 μmol·L^-1 CUDC-101 group.Cell scratch assay and Transwell chamber assay were used to detect the scratch area of the cells and the number of migration cells in the PCa DU145 cells before and after treated with CUDC-101； immunofluorescence staining was used to detect the expressions of epithelial cell marker E-cadherin （E-cadherin） and phosphorylated histone γ-H2AX in the PCa DU145 cells after treated with CUDC-101； Western blotting method was used to detect the expression levels of EMT-related protein， γ-H2AX， and phosphorylated protein kinase B （p-AKT） in the PCa DU145 cells after treated with CUDC-101. Results The TCGA Database and UALCAN Database analysis results showed that H2AX mRNA was highly expressed in the PCa tissue， and the disease-free survival （DFS） of the patients with low expression of H2AX was longer than those patients with high expression of H2AX mRNA （P<0.001）； the immunohistochemistry results showed that compared with adjacent benign prostate tissue，the rate of strong positive expression of H2AX protein in PCa tissue was increased （64.34% vs 14.29%）， and its over-expression was associated with the T stage of PCa （P=0.001） and World Health Organization（WHO）/International Society of Urological Pathology（ISUP） prognostic grapde group（GG） （P=0.004）， but was not associated with the patients’ age， Gleason score， lymphnode metastasis， or nerve and vascular invasion （P>0.05）； the immunofluorescence staining results showed that the compared with control and EMT induction groups，the fluorescence expressions of E-cadherin and γ-H2AX proteins in CUDC-101 treatment group were increased； the cell scratch assay and Transwell chamber assay results showed that compared with control group， the scratch healing area and number of migration DU145 cells in CUDC-101 treatment group was significantly decreased； the Western blotting results showed that compared with EMT induction group， the expression levels of E-cadherin and γ-H2AX proteins in the cells in CUDC-101 treatment group were increased （P<0.05 or P<0.01）， while the expression levels of Vimentin and p-AKT proteins were decreased （P<0.05 or P<0.01）. Conclusion Over-expression of H2AX protein is closely associated with poor prognosis of the patients with PCa. The novel HDACi inhibitor CUDC-101 can regulate the phosphorylations of H2AX and AKT and inhibit the EMT process in the PCa cells.

Objective To discuss the differentially expressed genes in severe bronchial asthma ［severe asthma （SA）］ by bioinformatics methods and analyze their mechanisms， and to screen the traditional Chinese medicines and their active components with potential therapeutic effects. Methods The GSE136587 and GSE158752 datasets were selected from the Gene Expression Omnibus （GEO） Database； R software was used for the differential analysis to obtain the differentially expressed gene； the protein-protein interaction （PPI） network analysis was used to screen the core genes， and the key pathways and hub genes were identified. The core genes were uploaded to the Coremine Database to screen for the traditional Chinese medicines with the potential therapeutic effects， and the relevant Chinese herbal prescriptions were searched in Chinese Medical Dictionary. Results A total of 466 differentially expressed genes were screened. The PPI network constructed through the STRING platform led to the selection of synaptosomal associated protein 25 kDa （SNAP25）， glutamate ionotropic receptor AMPA type subunit 2 （GRIA2）， neurexin 1 （NRXN1）， potassium voltage-gated channel subfamily a member 1 （KCNA1）， synaptotagmin 1 （SYT1）， and chromogranin A （CHGA）. The Gene Ontology （GO） functional enrichment analysis results showed that the biological processes of SA were significantly related to the cellular chemotaxis and leukocyte migration. The Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathways enrichment mainly involve bone marrow leukocyte migration，leukocyte chemotaxis，cell chemotaxis，leukocyte migaration， up-regulation of outside stimulus， and bone marrow leukocyte activation signaling pathways. Network pharmacology was applied to screen for 367 traditional Chinese medicines with potential therapeutic effects based on the core targets. Among them， ginseng， water buffalo horn， scorpio， and astragalus， which involve multiple core targets， were highly related to SA. A total of 17 potential Chinese herbal prescriptions with therapeutic effects were retrieved from Chinese Medical Dictionary. Conclusion The bioinformatics screening of potential biomarkers and traditional Chinese medicines with therapeutic effects for SA provides the new targets for the early diagnosis and research on the pathogenesis of SA， and offers new insights into the development of herbal prescriptions for its treatment.

Objective To discuss the differences in drug resistance， molecular typing， and biofilm-forming abilities of methicillin-resistant Staphylococcus aureus （MRSA） isolated from respiratory and non-respiratory tract specimens. Methods A total of 100 MRSA strains were selected from clinical specimens of the hospitalized patients， and 50 MRSA strains were isolated from the respiratory specimens and another 50 MRSA strains were isolated from non-respiratory tract specimens. These 100 MRSA strains underwent an in vitro susceptibility test for 14 kinds of antimicrobial drugs， molecular typing was performed through multilocus sequence typing （MLST） and Staphylococcus aureus protein A （Spa） typing， and their biofilm-forming abilities were detected by crystal violet staining assay. Results The in vitro susceptibility tests results showed that all the 100 MRSA strains had high overall resistance rates of over 60.0% to penicillin， erythromycin， clindamycin， levofloxacin， ciprofloxacin， moxifloxacin， and tetracycline， while only 9.0% exhibited resistance to trimethoprim-sulfamethoxazole， and all the strains were susceptible to vancomycin， linezolid， tigecycline， and quinupristin/dalfopristin. Compared with those isolated from non-respiratory tract specimens，the MRSA strains isolated from respiratory tract specimens showed significantly higher resistance rates to moxifloxacin， levofloxacin， ciprofloxacin， tetracycline， and gentamicin （P<0.05）. Among the 100 MRSA strains， 17 genotypes were identified， and the dominant types weve ST5-t2460 （26.0%）， ST239-t030 （23.0%）， and ST59-t437 （20.0%）. In the MRSA from the respiratory tract specimens， the dominant types were ST5-t2460 （20.0%） and ST239-t030 （13.0%）， followed by ST59-t437 （7.0%）； in the non-respiratory tract specimens， 13 genotypes were found， and the dominant types were ST59-t437 （13.0%） and ST239-t030 （10.0%）. All 100 MRSA strains were biofilm producers， among them the strong biofilm formers， moderate biofilm formers， and weak biofilm formers accounted for 2.0% （2/100）， 24.0% （24/100）， and 74.0% （74/100）， respectively. The ST59-t437 clones had overall strong biofilm-forming abilities， and 60.0% （12/20） stains were moderate to strong biofilm formers. Conclusion The MRSA strains isolated from the respiratory tract specimens have a significantly higher overall resistance rate than those isolated from the non-respiratory tract specimens. The ST59-t437 and ST239-t030 genotypes are dominant clones common to both types of specimens， and ST59-t437 strains show strong biofilm-forming abilities， and ST239-t030 strains exhibit the strongest resistance.

Objective To discuss the effect of forkhead box O1 （FOXO1） gene on the autophagy and apoptosis of the vascular smooth muscle cells in abdominal aortic aneurysm （AAA）， and to clarify its possible mechanism. Methods ^{The aneurysm tissue of nineteen AAA patients （AAA group） and adjacent normal aortic tissue of nineteen AAA patients （control group） were collected. Real-time fluorenscence quantitative PCR （RT-qPCR） method was used to detect the expression level of FOXO1 mRNA in aneurysm tissue of the subjects in two groups； transmission electron microscope was used to observe the autophagolysosome formation in aneurysm tissue of the subjects in two groups； Western blotting method was used to detect the expression levels of FOXO1 and autophagy-related proteins B cell lymphoma-2（Bcl-2）-binding protein（Beclin1）， microtubule-associated protein 1 light chain 3α （LC3）， and P62 proteins in aneurysm tissue of the subjects in two groups. The human aortic vascular smooth muscle cells （hVSMCs） were cultured in vitro and infected with FOXO1 siRNA （si-FOXO1） and its negative control （si-NC） lentivirus， then treated with 10 μmol·L－1 angiotensin Ⅱ （Ang Ⅱ） combined with autophagy activator rapamycin （Rap）. The cells were divided into blank control group， Ang Ⅱ group， Ang Ⅱ+ si-NC group， Ang Ⅱ + si-FOXO1 group， Ang Ⅱ+ si-NC + Rap group， and Ang Ⅱ+ si-FOXO1 + Rap group.CCK-8 assay was used to detect the proliferation activities of the cells in various groups； flow cytometry was used to detect the apoptotic rates of the cells in various groups； ELISA method was used to detect the levels of matrix metalloproteinase-2 （MMP-2） and matrix metalloproteinase-9 （MMP-9） in the cell supernatant in various groups； RT-qPCR method was used to detect the expression level of FOXO1 mRNA in the cells in various groups； Western blotting method was used to detect the expression levels of FOXO1， Bcl-2， Bcl-2 associated X prorein（Bax）， Cleaved-cysteinyl aspartate specific proteinase-3 （Cleaved caspase-3）， Beclin1， LC3， and P62 proteins in the cells in various groups. Results Compared with control group， the expression level of FOXO1 mRNA in aneurysm tissue of the subjects in AAA group was increased （P<0.05）， the number of autophagolysosomes was increased （P<0.05）， the levels of Beclin1 protein and the ratio of LC3 Ⅱ/LC3 Ⅰ were increased （P<0.05）， and the expression level of P62 protein was decreased （P<0.05）. Compared with blank control group， the proliferation activity of the hVSMCs in Ang Ⅱ group was decreased （P<0.05）， the apoptotic rate of the cells was increased （P<0.05）， the levels of MMP-2 and MMP-9 in the cell supernatant were increased （P<0.05）， the expression levels of Bax， Cleaved caspase-3， and Beclin1 proteins in the cells and the ratio of LC3 Ⅱ/LC3 Ⅰ were increased （P<0.05）， and the expression levels of Bcl-2 and P62 proteins were decreased （P<0.05）. Compared with Ang Ⅱ+ si-NC group， the proliferation activity of the hVSMCs in Ang Ⅱ+ si-FOXO1 group was increased （P<0.05）， the apoptotic rate of the cells was decreased （P<0.05）， the levels of MMP-2 and MMP-9 in the cell supernatant were decreased （P<0.05）， the expression levels of Bax， cleaved caspase-3， and Beclin1 proteins in the cells and the ratio of LC3 Ⅱ/LC3 Ⅰ were decreased （P<0.05）， and the expression levels of Bcl-2 and P62 proteins were increased （P<0.05）. Compared with Ang Ⅱ+ si-FOXO1 group， the apoptotic rate of the hVSMCs in Ang Ⅱ+ si-FOXO1 + Rap group was increased （P<0.05）， the levels of MMP-2 and MMP-9 in the cell supernatant were increased （P<0.05）， the expression level of Beclin1 protein in the cells and the ratio of LC3 Ⅱ/LC3 Ⅰ were decreased （P<0.05）， and the expression level of P62 protein was increased （P<0.05）. Conclusion Silencing the FOXO1 gene may increase the proliferation activity of the hVSMCs exposed to Ang Ⅱ by reducing the autophagy level and inhibiting the apoptosis， thus participating in the pathogenesis of AAA.}

Objective To analyze the expressions of glioma-associated oncogene homolog 1 （Gli1） and sex determining region Y 2 （Sox2） in the cervical cancer cells and their pathological significances， and to discuss the correlation between the expression of Gli1 and the stemness characteristics of the cervical cancer. Methods Immunohistochemical staining was used to detect the expression of Gli1， hypoxia-inducible factor 1-alpha （HIF-1α）， Sox2， sex determining region Y 9 （Sox9）， cluster of differentiation 44 （CD44）， octamer-binding transcription factor 4 （OCT4）， and lysine-specific demethylase 1 （LSD1） in 159 cases of cervical cancer tissues and their correlations were analyzed. Gene Expression Profiling Interactive Analysis （GEPIA） was used to clarify the effect of Gli1 expression on the prognosis of the patients with cervical carcinoma and its correlation with the stem cell markers. The sphere-forming ability and the expression of stem cell markers in the SiHa cells under hypoxic status were detected. Results The immunohistochemical staining results showed that the expression rate of Gli1 in cervical cancer tissue was positively correlated with the expression level of HIF-1α （r=0.374， P<0.001） and the expression level of Sox2 （r=0.176， P<0.05）. The GEPIA Database analysis results showed that the cervical cancer patients with positive Gli1 expression had poor prognosis （P<0.05）. Under hypoxic status， the expression levels of Gli1， HIF-1α， and Sox2 mRNA and proteins in the SiHa cells were significantly increased （P<0.001）， and the cells exhibited a strong sphere-formation ability. Conclusion Hypoxia upregulates the expression levels of Gli1 and Sox2 mRNA and protein in the cervical cancer cells，and promotes the formation of stemness characteristics of the cancer cells.

Objective To detect the stiffness of the gastrocnemius muscle in the patients with type 2 diabetes under different functional states by shear wave elastography （SWE） method， and to provide the objective basis for the early clinical detection of skeletal muscle damage. Methods A total of 70 patients with type 2 diabetes were selected as diabetes group， and 70 healthy examinees underwent medical check-ups were regared as control group. SWE was used to detect the Young’s modulus （E） value of the gastrocnemius muscle of the subjects in both groups under three different conditions（neutral ankle position at rest， plantar flexion ankle position during isometric contraction， and upright position during isometric contraction；the E values were normalized by body mass index （BMI） （E_BMI=E/BMI）； the differences in passive and active stiffness of the gastrocnemius muscle under various conditions of the subjects between two groups were compared； Pearson correlation analysis was used to assess the correlation of the E value in the diabetic patients with patients’ age， disease duration， level of glycated hemoglobin （HbA1c）， and level of advanced glycation end-products （AGEs）. Results There were no significant differences in the passive stiffness E values and E_BMI values of the gastrocnemius muscle of the subjects between two groups under neutral position （P>0.05）； there was no significant difference in the active stiffness E values of the gastrocnemius muscle under plantar flexion ankle position（P>0.05）， but the E_BMI value of the subjects in diabetes group were lower than that in control group （P<0.01）. Both E value and E_BMI value of active stiffness under upright position of the subjects in diabetes group were lower than those in control group （P<0.01）. The E value of active stiffness of the gastrocnemius of the subjects in diabetes group showed a negative correlation with disease duration， HbA1c level， and AGE level （r=－0.645， P<0.05； r=－0.741， P<0.05； r=－0.675， P<0.05）， and had no correlation with age （r=－0.116， P>0.05）. Conclusion The patients with type 2 diabetes exhibit reduced active stiffness in the gastrocnemius muscle， while passive stiffness may not be affected in the early stage. The use of SWE to measure active stiffness of the gastrocnemius muscle in standing isometric contraction can aid in the detection of subclinical muscle contraction function decline in the patients with type 2 diabetes.

Objective To discuss the effect of general transcription factor 2I （GTF2I） on the chemotherapeutic resistance to temozolomide in the glioblastoma multiforme （GBM）， and to clarify its mechanism. Methods Bioinformatics analysis was used to identify the common transcription factors for DNA methyltransferase 1 （DNMT1）， damage-specific DNA binding protein 1 （DDB1）， chromobox protein homolog 5 （CBX5）， and xeroderma pigmentosum complementation group C （XPC） in GBM tissue using the transcription factor prediction website （PROMO website）； correlation analysis and survival analysis for DDB1， CBX5， XPC， and DNMT1 with GTF2I and methylguanine methyltransferase （MGMT） were conducted based on The Cancer Genome Atlas （TCGA） Database；small interfering RNAs （siRNAs） were used to transfect and silence the gene expression of MGMT and GTF2I in the human T98 GBM cells and LN229 glioma cells； real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression level of the gene mRNA. After silencing the GTF2I gene， plate clone formation assay was used to detect the colon forming ability of the tumor cells， and CCK-8 assay was used to detect the sensitivity of the cells to temozolomide. Results The bioinformatics analysis results showed that the expression levels of DDB1， CBX5， XPC， and DNMT1 were significantly positively correlated with the expression level of GTF2I in GBM tissue （P<0.05）， and were negatively correlated with the expression level of MGMT （P<0.05）. The expression level of GTF2I was significantly negatively correlated with the expression level of MGMT （P<0.05）. Excluding the GBM patients who did not receive temozolomide treatment， the survival analysis results indicated that the patients with high expression of GTF2I had a decreased overall survival time. After silencing the MGMT gene， the expression levels of GTF2I， DDB1， CBX5， and XPC mRNA in the human brain glioma T98 cells were increased （P<0.001）； after silencing the GTF2I gene， the expression level of MGMT mRNA in the human brain glioma LN229 cells was increased （P<0.05）， while the expression levels of DDB1， CBX5， XPC， and DNMT1 mRNA were significantly decreased （P<0.05 or P<0.001）. The plate clone formation assay results showed that there was no significant difference in the clone formation ability of the cells before and after silencing the GTF2I gene （P=0.138）. The CCK-8 assay results showed that compared with control group，the viability of the cells in observation group was decreased （P<0.05）. Conclusion The transcription factor GTF2I regulates the expressions of key DNA damage repair protein mRNA， including MGMT， DDB1， CBX5， and XPC， and is involved in the chemotherapeutic resistance of the GBM cells to temozolomide；GTF2I may represent a potential new therapeutic target for the GBM.

Objective To study the clinical efficacy of endoscopic-assisted subgingival scaling combined with erythritol subgingival sand blasting technology for the treatment of peri-implantitis，and to provide the theoretical basis for the effective treatment of peri-implantitis. Methods The patients with peri-implantitis who attended the Periodontology Department in our hospital and received treatment were selected. A total of 58 peri-implantitis patients were sequentially observed according to the time of visit and were randomly divided into control group （28 cases） and minimally invasive group （30 cases）； the patients in control group underwent traditional blind subgingival curettage， while the patients in minimally invasive group were received endoscopic-assisted subgingival scaling combined with erythritol subgingival sandblasting. The probing depth （PD）， modified plaque index （mPLI）， modified sulcus bleeding index （mSBI）， and levels of interleukin-1β （IL-1β）， interleukin-6 （IL-6）， and tumor necrosis factor-α （TNF-α） in gingival crevicular fluid of the patients in two groups were analyzed before and after treatment. Results Before treatment， there were no significant differences in PD， mPLI， mSBI， and levels of IL-1β， IL-6， and TNF-α in gingival crevicular fluid of the patients between two groups （P>0.05）， demonstrating there was comparability. Compared with before treatment， the PD， mPLI， mSBI， and levels of IL-1β， IL-6， and TNF-α in gingival crevicular fluid of the patients in two groups after treatment were decreased （P<0.05）.Compared with control group， the PD， mPLI， and mSBI of the patients in minimally invasive group were obviously decreased （P<0.05）， and the levels of IL-1β， IL-6， and TNF-α were also significantly decreased （P<0.05）. Conclusion In the short term， endoscopic-assisted subgingival scaling combined with erythritol subgingival sandblasting technology for the treatment of peri-implantitis is more beneficial in controlling inflammation around the peri-implant tissue and improving clinical symptoms.

Objective To discuss the association between hemodynamic patterns and the risk of ischemic stroke in the people with chronic orthostatic intolerance （OI）， and to provide the basis for the early assessment of the onset of ischemic stroke. Methods A total of 638 residents aged ≥40 years from three streets/townships in Changchun City of Jilin Province were selected as the subjects by multistage random sampling method. Transcranial Doppler （TCD） combined with head-up tilt test （HUTT） were used to evaluate the hemodynamic patterns of the people with OI， and the OI research cohort was established. The patients were divided into orthostatic hypotension （OH） group， orthostatic hypertension （OHT） group， postural tachycardia syndrome （POTS） group， and orthostatic cerebral hypoperfusion syndrome （OCHOs） group according to the changes of hemodynamics. The subjects were followed up once every six months for two years， and the first incidence of ischemic stroke of the subjects was regarded as the observation endpoint；the baseline data and incidence of ischemic stroke of the subjects during the follow-up period were collected； Cox proportional hazards model was used to analyze the association factors with the risk of ischemic stroke. Results A total of 121 subjects were diagnosed with OI， including 80 subjects （66.12%） in OH group， 35 subjects （28.93%） in OHT group， 5 （4.13%） subjects in POTS group， and 1 subject （0.82%） in OCHOs group. During the follow-up period， the overall people had 43 confirmed new cases of ischemic stroke. After controlling the related confounding factors， compared with non-OI individuals， the risk of ischemic stroke in the entire OI people， as well as in the OH and OHT patients， was increased by 1.527 times ［hazard ratio （HR）=2.527， 95% confidence interval （CI）： 1.269－5.032， P<0.01）］， 2.268 times （HR=3.268， 95% CI： 1.603－6.663， P=0.001）， and 2.153 times （HR=3.153， 95% CI： 1.213－8.916， P=0.008）， respectively. Conclusion OI is associated with the increased risk of ischemic stroke， particularly in the patients in OH and OHT groups.

Objective To discuss the expression levels of mitochondrial autophagy and apoptosis-related genes in peripheral blood mononuclear cells （PBMCs） in the patients with myasthenia gravis （MG），and to clarify its diagnostic value for MG. Methods The PBMCs from 50 healthy controls （control group） and 50 MG patients （MG group） were collected and extracted. Real-time fluorescence quantitative PCR （RT-qPCR） and Western blotting methods were used to detect the expression levels of mitochondrial autophagy factors phosphatase and tensin homolog （PTEN）-induced kinase 1 （PINK1）， E3 ubiquitin-protein ligase PARK2 （Parkin）， ubiquitin-binding protein p62 （p62）， microtubule-associated proteins light chain 3 type Ⅱ （LC3Ⅱ）， and apoptotic factors cytochrome C （Cyt-C）， B cell lymphoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， cysteine-containing aspartic acid protease 3 （caspase 3）， and cysteine-containing aspartic acid protease 9 （caspase 9） mRNA and protein. Different cut-off values were set based on the mRNA detection results， and the corresponding sensitivity and specificity at each cut-off value were calculated. The receiver operating characteristic （ROC） curve was drawn. Results Compared with control group， the expression levels of autophagy factors PINK1， Parkin， and LC3Ⅱ mRNA and protein in the PBMCs in MG group were significantly decreased （P<0.01）， while the expression levels of p62 mRNA and protein were increased （P<0.05 or P<0.01）. The expression levels of apoptotic factors Cyt-C， Bax， caspase 3， and caspase 9 mRNA and protein were significantly increased （P<0.05 or P<0.01）， and while the expression levels of Cyt-C mRNA and protein were increased （P<0.05 or P<0.01）， and the expression levels of Bcl-2 mRNA and protein were significantly decreased （P<0.01）. The ROC curve analysis results showed that the area under the curve （AUC） for PINK1， Parkin， p62， and LC3Ⅱ were 0.969 6 （95%CI： 0.943 5-0.995 7）， 0.944 0 （95%CI： 0.904 7-0.983 3）， 0.855 6 （95%CI： 0.776 7-0.934 5）， and 0.908 8 （95%CI： 0.852 6-0.965 0） （P<0.01）， respectively，and the sensitivities were 92%， 92%， 78%， and 84%， and the specificities were 88%， 82%， 92%， and 82%， respectively； whereas the AUC for Cyt-C， Bax， Bcl-2， caspase 3， and caspase 9 was 1.000 （P<0.01）， and the sensitivity and specificity reached 100%. Conclusion Mitochondrial autophagy disorder and increased apoptosis are found in the PBMCs in the patients with MG， which has a higher guiding value for clinical diagnosis of MG.

Objective To discuss the genotype distribution of methicillin-resistant Staphylococcus aureus （MRSA） in a tertiary hospital and to discuss the correlation among different molecular types of the strains， and to construct the resistance profile model under different genetic backgrounds. Methods A total of 204 strains of Staphylococcus aureus （S. aureus）from 25 departments of the hospital were selected. The automatic VITEK 2 Compact system and E-test strips were used to detect the antimicrobial susceptibility of the strains. The detection of mecA gene by the polymerase chain reaction （PCR） method was used as the confirmatory experiment， the abilities of cefoxitin （FOX） and（or） oxacillin （OXA） were compared as phenotypic detection methods to screen MRSA. The molecular typing of MRSA strains was carried out by PCR method， including Staphylococcal proteinA gene （spa） determination， accessory gene regulator （agr） typing， multilocus sequence typing （MLST）， and staphylococcal chromosomal cassettes mec （SCCmec） typing. The resistance profile was constructed combing with antimicrobial susceptibility tests results and molecular typing results. Results A total of 39 MRSA strains were obtained by detecting the mecA gene. A total of 51 phenotypic MRSA strains were identified by testing FOX and OXA. In spa typing， 57 different types were identified， including 5 new types （t20226， t20227， t20228， t20229， and t20230）， with the main types being t309 （30.9%）， t078 （11.8%）， and t437 （11.8%）. In agr typing， 94.9% of MRSA belonged to agr Ⅰ. The MLST analysis results of MRSA populations showed that ST59 clone （61.5%） was the most prevalent， followed by ST72 （20.5%）. A total of 87.2% of MRSA carried type Ⅳ SCCmec， with subtypes Ⅳa accounting for 24 strains and subtype ⅣF accounting for 10 strains. Conclusion The main genotype of MRSA is ST59-t437-agr Ⅰ-Ⅳa and its resistance profile is primarily characterized by resistance to FOX-OXA-penicillin （PEN）-erythromycin （ERY）-clindamycin （CLI）.

Objective To discuss the expression levels of interleukin-33 （IL-33）， interleukin-8 （IL-8）， and the major components of neutrophil extracellular traps （NETs）-myeloperoxidase （MPO） and citrullinated histone H3 （CitH3） in lung tissue of the patients with chronic obstructive pulmonary disease （COPD）， and to clarify the effects of IL-33， IL-8， and NETs on the occurrence and development of COPD. Methods Seventy-seven patients underwent lobectomy due to pulmonary nodules or lung tumors were selected as the subjects and divided into non-smoking control group （20 cases）， smoking control group （18 cases）， non-smoking COPD group （19 cases）， and smoking COPD group （20 cases）.The lung tissue samples from the subjects were collected.HE staining was used to observe the pathomorphology of lung tissue of the patients in various groups；immunohistochemisty staining was used to detect the expression levels of IL-33， IL-8， CitH3， and MPO proteins in lung tissue of the patients in various groups； Spearman correlation analysis was used to detect the relationship between expression levels of IL-33， IL-8， CitH3， and MPO proteins in lung tissue of the patients in COPD groups and their correlations with lung function index， mean linear intercept （MLI）， mean alveolar number （MAN）， and Bosken score. Results Compared with non-smoking control group， the airway Bosken score of the patients in smoking control group was significantly increased （P<0.001）； compared with non-smoking control and smoking control groups， the airway Bosken score and MLI of the patients in non-smoking COPD group were significantly increased （P<0.05）， while the MAN was significantly decreased （P<0.05）； compared with non-smoking control group， smoking control group， and non-smoking COPD group， the airway Bosken score of the patients in smoking COPD group was significantly increased （P<0.05）； compared with non-smoking control group and smoking control group， the MLI of the patients in smoking COPD group was significantly increased （P<0.05）， and the MAN was significantly decreased （P<0.05）. The immunohistochemisty staining results showed that compared with non-smoking control group， smoking control group， and non-smoking COPD group，the expression level of IL-33 in lung tissue of the patients in smoking COPD group was significantly increased （P<0.05）； compared with non-smoking control group and smoking control group，the expression levels of IL-8， CitH3， and MPO proteins in lung tissue of the patients in smoking COPD group were significantly increased （P<0.05）. Compared with non-smoking control group and smoking control group， the expression levels of IL-33， IL-8， CitH3， and MPO proteins in lung tissue of the patients in non-smoking COPD group were significantly increased （P<0.05）.The Spearman correlation analysis results showed that there was a negative correlation between the expression level of IL-33 protein and forced expiratory volume in first second/forced vital capacity （FEV1/FVC）， the forced expiratory volume in first second predicted FEV1 （FEV1%pred）， and MAN （r=－0.406， P<0.05； r=－0.493， P<0.01； r=－0.567， P<0.05） in lung tissue of the patients in COPD group， and there was a positive correlation with Bosken score and MLI （r=0.935， P<0.001； r=0.590， P<0.001）；the expression level of IL-8 protein in lung tissue of the patients in COPD group was negatively correlated with the FEV1/FVC， FEV1%pred， and MAN （r=－0.527， P<0.01； r=－0.497， P<0.01； r=－0.463， P<0.01）， and was positively correlated with the Bosken score and MLI （r=0.557， P<0.001； r=0.486， P<0.01）； the expression level of CitH3 protein in lung tissue of the patients in COPD group was negatively correlated with the FEV1/FVC， FEV1%pred， and MAN （r=－0.527， P<0.01； r=－0.640， P<0.001； r=－0.531， P<0.01）， and was positively correlated with the Bosken score and MLI （r=0.565， P<0.001； r=0.585， P<0.001）；the expression level of MPO protein in lung tissue of the patients in COPD group was positively correlated with the FEV1/FVC （r=－0.329， P<0.05）， and was positively correlated with the Bosken score （r=0.410， P<0.05）； the expression level of IL-8 protein was positively correlated with the expression levels of CitH3 and MPO proteins （r=0.390，P<0.05； r=0.349， P<0.05）； the expression level of IL-33 protein was positively correlated with the expression levels of IL-8， CitH3， and MPO proteins （r=0.602，P<0.001； r=0.616，P<0.001； r=0.387， P<0.05）. Conclusion The expression levels of IL-33， IL-8， and NETs in lung tissue of the COPD patients are increased， and they may be involved in the chronic inflammation of COPD and correlated with the severity of the disease.

Objective To compare the differences in noninvasive perfusion index （PI） before and after extubation between the right palm （ preductal） and right sole （postductal） in the preterm infants，and to discuss whether the conventional frequency mechanical ventilation （MV） mode has an effect on it， and to clarify the correlation between the respiratory severity score （RSS） and PI. Methods The preterm infants with the gestational age （GA） ≥32 weeks and body weight （BW） ≥1 500 g who were born with respiratory distress and underwent noninvasive assisted ventilation therapy， with pulse oxygen saturation （SpO₂） <90%， requiring conventional frequency MV assistance， were selected. A total of 55 patients met the inclusion criteria were included. The extubation was carried out after meeting the criteria 24 h after birth. The median values within 30 s after stabilization of PI in the right palm and right sole were recorded before and after extubation. At the same time， the ventilator parameters such as the fraction of inspired oxygen （FiO₂） and the mean airway pressure （P_mean） before extubation were recorded. The paired samples t-test was used to compare the differences in PI of the right palm and right sole before and after extubation；multivariate linear regression analysis was used to analyze the correlation between GA， BW， and RSS with the preductal PI of the right palm before extubation and the correlation between FiO₂ and P_mean with PI. Results The PI of the right palm before extubation was lower than after extubation （P<0.05）； the PI of the right sole before extubation was lower than that after extubation （P<0.05）； the linear regression analysis results showed there was no correlation between GA and PI （P>0.05）， there was a positive correlation between BW and PI ［b=0.44，standardized regression coefficient （β）=0.25， P<0.05）］， and there was a negative correlation between RSS and PI （b=－0.56， β=－0.68， P<0.05）， and the regression equation was PI=1.9+0.44×BW－0.56×RSS；the further multivariate linear regression analysis results of the ventilator parameters showed that the ventilator parameters constituting the RSS， FiO₂ （b=－2.52， β=－0.27， P<0.05） and P_mean （b=－0.39， β=－0.63， P<0.05）， both showed a linear relationship with PI and they were risk factors for it， and the β value of P_mean was greater than that of FiO₂， indicating that the former had a greater impact on PI. Conclusion The conventional frequency MV mode can affect PI， and RSS under this mode is a risk factor for PI； higher RSS can have an adverse effect on the circulation， and P_mean has a greater impact on PI compared with FiO₂.

Objective To discuss the effect of microRNA-151 （miR-151） on the biological behavior of the human trophoblast cells HTR-8/SVneo under hypoxic conditions，and to clarify the potential mechanism. Methods A total of 47 parturients with preeclampsia （preeclampsia group） and 36 parturients （ normal group） were selected as the research subjects. The expression level of miR-151 in placenta tissue of the subjects in two groups was detected by real-time fluorescence quantitative PCR （RT-qPCR） method. The miR-151 inhibitor and its negative control inhibitor NC were transfected into the HTR-8/SVneo cells， followed by exposure to hypoxia （1% O₂） for 48 h to establish control， hypoxia， hypoxia + inhibitor NC， and hypoxia + inhibitor groups. RT-qPCR method was used to detect the expression levels of miR-151 in the cells in various groups； MTT assay was used to detect the survival rates of the cells in various groups； Transwell chamber assay was used to detect the migration and invasion numbers of the cells in various groups； Western blotting method was used to detect the expression levels of matrix metalloproteinases （MMP）-2 and MMP-9， and epithelial-mesenchymal transition （EMT） related proteins in the cells in various groups； Bioinformatics analysis was used to predict the downstream target genes of miR-151， and the intersection target genes were further analyzed for protein-protein interaction （PPI） network by STRING Database. Results Compared with normal group， the expression level of miR-151 in placenta tissue of the patients in preeclampsia group was significantly increased （P<0.05）. Compared with control group， the proliferation activity， number of invasion cells， and number of migration cells of HTR-8/SVneo cells in hypoxia group were significantly decreased （P<0.05）， the expression levels of MMP-2， MMP-9， N-cadherin， and vimentin in the cells were significantly decreased （P<0.05）， and the expression levels of miR-151 and E-cadherin were significantly increased （P<0.05）. Compared with hypoxia group， the expression levels of MMP-2， MMP-9， N-cadherin， and vimentin in the cells in hypoxia + inhibitor group were significantly increased （P<0.05）； the levels of miR-151 and E-cadherin were significantly decreased （P<0.05）； while there were no significant differences in the above indexes in hypoxia + inhibitor NC group （P>0.05）. The bioinformatics analysis results showed that 34 potential target genes of miR-151， among which RCCCH-type zinc finger protein 1 （RC3H1）， AGO2， AGO3， Fragile X related protein 1 （FXR1）， and transformer 2β （TRA2B） may be the key potential target genes. Conclusion miR-151 is highly expressed in placenta tissue of the patients with preeclampsia. The downregulation of miR-151 expression can promote the proliferation， invasion， and migration of the trophoblast cells under hypoxic conditions.

Objective To evaluate the diagnostic value of non-invasive models for autoimmune cirrhosis with esophagogastric varices （EGV），and to provide the basis for the early diagnosis of autoimmune cirrhosis with EGV. Methods The retrospective collection of clinical data from 238 patients diagnosed with autoimmune cirrhosis was performed， and the patients divided into EGV group and non-EGV group according to whether complicated with EGV. The levels of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， γ-glutamyltransferase （γ-GT）， platelet （PLT）， AST / PLT index （APRI）， fibrosis-4 index （FIB-4）， and AST/ALT ratio （AAR） of the patients in both groups were compared and the receiver operating characteristic （ROC） curve was plotted to calculate the area under the curve （AUC）， sensitivity， specificity， positive predictive value， and negative predictive value to evaluate the diagnostic values of various models for autoimmune cirrhosis with EGV. Results The levels of APRI， FIB-4， and ALT of the patients in EGV group were significantly higher than those in non-EGV group （P<0.01）， while the levels of PLT and AAR were significantly lower than those in non-EGV group （P<0.01）. In single indicator diagnostic model， ALT had the largest AUC of 0.645 （95% CI： 0.580－0.705， P<0.001）， the sensitivity was 93.75%， the specificity was 34.04%， the positive predictive value was 68.53%， and the negative predictive value was 78.05%. In multiple indicator combined models， the combined model of ALT+FIB-4+AAR+PLT had the largest AUC of 0.787 （95% CI： 0.730－0.838， P<0.001）， and the sensitivity was72.22%， the specificity was 73.40%， the positive predictive value was 80.62%， and the negative predictive value was 63.30%. Compared with ALT and ALT+FIB-4 models，the AUC of the combined models of ALT+FIB-4+AAR， ALT+FIB-4+AAR+PLT， and ALT+FIB-4+AAR+PLT+APRI had statistically significant differences （P<0.05）. However， there were no significant differences in AUC among the above three models （P>0.05）. Conclusion The combined detection of APRI， FIB-4， ALT， PLT， and AAR can improve the early diagnostic efficacy of autoimmune cirrhosis with EGV.

Objective To report the diagnosis and treatment of one patient with acute left lower limb arterial embolism complicated with myonephropathic metabolic syndrome （MNMS）-associated heart failure， and to provide the reference for the diagnosis， anesthesia， and treatment of such patients. Methods The retrospective analysis on the clinical data， anesthesia methods， and perioperative management of one patient with acute left lower limb arterial embolism complicated with MNMS-associated heart failure was conducted， and the related literatures were reviewed. Results The patient， a 56-year-old male， was admitted to the hospital on March 25， 2017， due to sudden pain and numbness in the left lower limb accompanied by sensory and motor disturbances. The vascular ultrasound performed 35 min after the admission suggested a secondary thrombus from the distal superficial femoral artery to the entire popliteal artery. The laboratory tests performed 77 min after admission showed the myoglobin （MB） level was 698.7 μg·L^－1， cardiac troponin I （TnI） was 0.092 μg·L^－1， and creatine kinase isoenzyme （CK-MB） was 4.78 μg·L^－1. The electrocardiogram results taken 63 min after admission indicated tachycardia， left ventricular hypertrophy， atrial fibrillation， and atrioventricular block. The initial diagnosis was acute left lower limb arterial thrombosis， coronary artery disease， old myocardial infarction， arrhythmia， and type 2 diabetes. The patient was scheduled for an emergency thrombectomy of the left lower limb under general anesthesia 3 h after admission. During the surgery， the patient’s vital signs were stable， but 8 min after operation， the patient suddenly developed ventricular fibrillation， which was considered to be a reinfarction of acute myocardial infarction complicated with MNMS. After active rescue treatment， the patient’s life was still not saved. Conclusion For the patients with acute limb arterial embolism complicated with heart failure， timely restoration of limb blood supply is crucial in treatment. Appropriate fluid resuscitation to expand blood volume， maintaining electrolyte balance， and protecting cardiac and renal functions can effectively reduce the mortality and amputation rate of the patients.

Objective To observe the clinical efficacy of autologous platelet-rich fibrin （PRF） application alone in immediate dental implant placement in the molars with periapical periodontitis， and to discuss the mechanism， in order to widen its clinical application and to provide the guidance for its clinical practice. Methods The clinical data of one patient who underwent immediate implant placement in the molars with periapical periodontitis using PRF as the sole material were collected. The changes in the tissues surrounding the implant were evaluated through the three-dimensional reconstruction of cone-beam CT （CBCT） and oral scan data， the therapeutic methods and outcomes of PRF treatment were analyzed combined with the relevant literatures. Results A minimally invasive extraction of the patient’s 46 diseased molar was conducted followed by the immediate implant placement. Before surgery， 30 mL of the patient’s own blood （3 tubes） was drawn， which was then placed in 10 mL glass-coated plastic tubes without anticoagulants. Three PRF were prepared with the blood by centrifuging at 3 000 r·min^－1 for 10 min. These clots were used as the only filling material for the jumping gap. The postoperative CBCT and oral scan 3D reconstruction results showed that the peri-implant bone tissue was increased by 203.19 mm³， the buccal bone height was increased by 5.83 mm， and the buccal bone tissue was increased by more than 1 mm at 6 months postoperatively； after 12 months， the soft and hard tissues around the implant remained essentially stable. Conclusion The application of autologous PRF alone in immediate implant placement in the molars with periapical periodontitis achieves favorable treatment outcomes， the peri-implant bone tissue regenerates， and the peri-implant hard and soft tissues remain stable， which providing the new insights into the immediate implant treatment.

Objective To analyze the clinical data of a rare case of gastrointestinal stromal tumors （GISTs） with multifocal heterochronous primary resistance due to temporal gene mutations after long-term treatment with imatinib， and to reveal the importance of secondary mutation heterogeneity in the recurrence and resistance process of GISTs. Methods The clinical data of one patient with recurrent GISTs were collected. The histomorphology observation was observed by HE staining；the expressions of related proteins were detected by immunohistochemistry；first and second generation sequencing techniques were used to analyze the tumor gene mutations. Results The patient， a 66-year-old male， underwent partial gastrectomy for gastric stromal tumor.The first-generation sequencing results identified it was a proto-oncogenea and there was a deletion mutation in exon 11 of the tyrosine protein kinase kit （KIT） gene （p.551-554del）. Approximately one year after imatinib treatment and a subsequent 4-month drug holiday， the patient experienced a relapse with a splenic lesion.The continuous first-line treatment with imatinib controlled the splenic lesion. After 59 months， a new pelvic mass was identified， and the patient received increased doses of imatinib （600 mg·day^－1） along with second-line therapy with sunitinib. After treated for two months，the CT scan results showed that the size of the splenic lesion tended to stabilize， while the volume of the new pelvic lesion continued to increase. The second generation sequencing results indicated that on the basis of the original KIT gene exon 11 deletion， subsequent mutations in exon 13 （p.V654A） and exon 17 （p.Y823D） of the KIT gene occurred separately in two recurrent lesions. Conclusion Under the pressure of targeted drug selection， the biological behavior of GISTs shows complexity，such as，time heterogeneity，spatial heterogeneity and molecular heterogeneity； different recurrent lesions may exhibit different resistance mechanisms.

Objective To analyze the clinical presentations， radiographic features， operative findings， and pathological characteristics of one patients with unilateral maxillary and mandibular simple bone cysts （SBC）， and to enhance the clinicians’ recognition and treatment of this condition. Methods The clinical data， radiographic features， operative findings， pathological characteristics， clinical diagnosis， and treatment of a case with right-sided maxillary and mandibular SBC were collected，and the relevant literatures were reviewed. Results The patient， an 11-year-old male， presented with a clearly demarcated low-density image within the right maxillary and mandibular bones on panoramic tomography. There was no history of trauma， no subjective symptoms or facial asymmetry before treatment， and no positive signs of the specialist examination. The patient was diagnosed with right-sided maxillary and mandibular SBC based on the operative exploration and pathological diagnosis. The patient received the conventional curettage treatment and recovered well postoperatively without significant discomfort. A 6-month follow-up results showed good intraoral wound healing without swelling. The cone beam computed tomography （CBCT） results showed a smaller bone cavity with visible new bone trabeculae， indicating good osteogenesis. The patient was currently the under regular follow-up. Conclusion Unilateral maxillary and mandibular SBC do not present with characteristic clinical signs and radiographic features， which can be easily confused with the common maxillofacial diseases. A definitive diagnosis should be made by combining radiographic examination with surgical and pathological findings.

Keratinized gingiva plays an important role in maintaining the health of the periodontal tissue，however， the specific mechanism of its formation is not fully understood. The differentiation of oral epithelial cells is determined by genetics，but the type of epithelium is also influenced by the underlying connective tissue and periodontal ligament，and the elastic fibers in the connective tissue has an important indirect effect on maintaining the epithelial phenotype. Due to the unclear formation mechanism， the primary treatment method for augmenting keratinized gingiva is currently mucogingival surgery，and the employing graft materials included autogenous tissues（free gingival grafts，subepithelial connective tissue grafts，and autologous platelet concentrates，etc），allogeneic materials （acellular dermal matrix，xenogeneic collagen matrix， and amniotic membrane，etc.） and the combination of these in the form of tissue-engineered products （using layered chitosan or nanofiber hydrogel composite materials as scaffolds）. This review covers the keratinization process of the gingiva，the factors determining keratinization，and the existing graft materials， summarizes the current research advancements in gingival keratinization mechanism and materials for gingival augmentation， and provides the basis for the further mechanism research and the development of new materials.

The dental mesenchymal stem cells （DMSCs） are mesenchymal stem cells derived from the neural crest ectoderm and have exceptional self-renewal and multilineage differentiation capabilities. The DMSCs are extensively used in tissue engineering and regenerative medicine research. The DMSCs can be expanded in vitro on a large scale to meet the needs of research and therapy by three-dimensional culture technique. Compared with traditional two-dimensional cell culture techniques，three-dimensional culture more effectively simulates the structure and microenvironment that the stem cells encounter in vivo，providing simultaneous spatial and temporal regulation of the proliferation and differentiation of the stem cells. Various three-dimensional in vitro culture techniques have been developed in recent years. Hanging drop culture is straightforward， but controlling the tissue culture environment is challenging； microfluidic chips offer better control over cellular parameters， but are costly and face the technical platform challenges，so their widespread application is limited； magnetic levitation culture is cost-effective and easy to perform with the rapid cell spheroid formation， but its magnetization effect make it unsuitable for the quantitative analysis. Other three-dimensional culture methods include rotating cell culture systems， centrifugation for spheroid culture， overlay culture and artificial scaffold methods，and they all have their own advantages and limitations. This review encompasses the different methods of three-dimensional culturing of DMSCs and their applications in various tissue regeneration and disease treatment，and provide the reference for the precise regulation of the function of the DMSCs and the research of regenerative medicine.

14-3-3ε protein， also known as YWHAE protein， is a member of the 14-3-3 protein family characterized by a small relative molecular weight. Unlike protein kinases， 14-3-3ε protein does not participate in protein phosphorylation within the cells； instead， it forms the unique dimer structures and binds to the specific phosphorylated proteins，thereby regulating the activity and subcellular localization of the protein ligands and playing a role in the regulation of various cellular activities. The aberrant expression of 14-3-3ε protein has been discovered in the numerous cancers. The altered regulatory effect of aberrantly expressed 14-3-3ε protein on its protein ligands can affect the ligands’ subcellular localization and enzymatic activity， leading to the development， invasion， and metastasis of the tumor cells. The dimer structure of 14-3-3ε protein is a critical structural basis for its biological functions in the tumor progression， and disrupting the dimer structure could be targeted to kill the tumor cells. This review is based on the domestic and international research findings on the 14-3-3ε protein and covers its structure，mechanism， and the impact and mechanisms during the development process of gastric cancer， liver cancer， skin cancer， and various other tumors.

Cyclic nucleotide-gated （CNG） ion channel is a type of non-selective tetrameric cation channel that can be directly activated by intracellular signaling molecules—cyclic nucleotides， and is one of the main pathways for calcium ions to enter the cells. The CNG channel proteins are encoded by six different genes： four A subunits and two B subunits. The activity of CNG ion channel can be modulated by calcium/calmodulin （Ca²⁺/CaM） as well as phosphorylation or the action of phosphatidylinositol on the membrane， thereby altering intracellular calcium ion concentrations and participating in the regulation of a variety of biological functions. Since the discovery of CNG ion channels in rod photoreceptors， it has undergone research processes such as physiological functions， cloning related genes， understanding regulatory mechanisms， analyzing crystal structures， and developing related gene therapy methods. They play an important role in signal transduction in olfactory sensory neurons （OSNs）of vision and smell. This article briefly reviews the function， structure， regulatory mechanism， and relationship with related diseases of CNG ion channel， in order to provide the theoretical basis and reference for the treatment of CNG ion channel-related diseases.