Objective To discuss the effect of salidroside on the apoptosis of the bone marrow CD71+ nucleated red blood cells（RBC） in the rat model of high altitude polycythemia （HAPC），and to clarify its mechanism. Methods orty-five male SD rats were randomly divided into control group （exposed to 1 500 m altitude + NaCl solution）， model group （exposed to 5 000 m altitude + NaCl solution）， and drug group （exposed to 5 000 m altitude + 0.15 g·kg^－1·d^－1 salidroside），and there were 15 rats in each group. The RBC counts， hemoglobin （Hb） levels， and hematocrit （HCT） levels of the rats in various groups were detected； flow cytometry was used to detect the percentages of bone marrow CD71+nucleated RBC， apoptotic rates of bone marrow CD71+nucleated RBC， levels of mitochondrial membrane potential （MMP） and expression levels of cysteinyl aspartate-specific protease-3 （Caspase-3） in bone marrow CD71+nucleated RBC of the rats in various groups； Western blotting method was used to detect the expression levels of B-cell lymphoma-2 （Bcl-2）， Bcl2-associated X protein （Bax） ，and cysteinyl aspartate-specific protease-9 （Caspase-9） proteins in bone marrow CD71+nucleated RBC of the rats in various groups. Results Compared with control group， the RBC count， Hb level， HCT level of the rats in model group were increased （P<0.01）， and the percentage and apoptotic rate of bone marrow CD71+nucleated RBC were increased （P<0.01）， and the expression levels of Bax， Caspase-9， and Caspase-3 proteins in the bone marrow CD71+nucleated RBC were increased （P<0.05）. Compared with model group， the percentage of bone marrow CD71+nucleated RBC of the rats in drug group was decreased （P<0.01）， the apoptotic rate of bone marrow CD71+nucleated RBC（P<0.01）， and the MMP level in the bone marrow CD71+nucleated RBC were increased （P<0.05）， and the expression levels of Caspase-3， Bax， and Caspase-9 proteins in the bone marrow CD71+nucleated RBC were decreased （P<0.01）. Conclusion Salidroside can effectively inhibit the increasing of apoptosis of nucleated RBC in bone marrow of the HAPC rats and improve the excessive accumulation of the RBC caused by hypoxia， which may have a certain preventive and protective effect in the pathogenesis of HAPC in the rats.

Objective To screen the active ingredients of traditional Chinese medicine Sanghuang in the treatment of pneumonia based on network pharmacology， and to analyze the active ingredients and targets of Sanghuang in the treatment of pneumonia. Methods The Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） software was used to search for the ingredients and targets of Sanghuang； the Gene Name was obtained from the Uniprot database； the targets corresponding to pneumonia were found through GeneCards， and the targets of Sanghuang and pneumonia were compared to obtain the key targets； Cytoscape 3.9.1 was used to draw the chemical ingredients of Sanghuang-pneumonia-target network diagram； the protein-protein interaction （PPI） network diagram was constructed by String platform； Gene Ontology （GO） biological function analysis and Kyoto Encyclopedia of Genes and Genomes（KEGG） signaling pathway enrichment analysis on key genes of Sanghuang were performed by using the ClueGo plugin in Cytoscape software. Results There were 21 kinds of active ingredients in the traditional Chinese medicine Sanghuang， and （E）-4-（3，4-dihydroxyphenyl）but-3-en-2-one contained the most targets， including cytochrome P450 family （CYP450）， human epidermal growth factor receptor （EGFR）， and matrix metalloproteinase （MMP） and so on. A total of 358 targets were screened as the compound targets of Sanghuang，among them 217 genes were regarded as common targets for Sanghuang and pneumonia， and 69 key genes related to pneumonia were amine oxidase copper containing 3 gene （AOC3）， DNA ligase 1 gene （LIG1）， glutamyl aminopeptidase gene （ENPEP）， aldehyde dehydrogenase 2 family member gene （ALDH2）， PHD finger protein 8 gene （PHF8）， solute carrier family 11 member 2 gene （SLC11A2），CYP50 and so on.The KEGG and GO analysis results showed that Sanghuang mainly produced a marked effect through 17 metabolic pathways， including the phosphatidylinositol-3 kinase/protein kinase B （PI3K/Akt） signaling pathway， microRNAs（miRNA） in cancer， chemical carcinogenesis - receptor activation， prostate cancer， proteoglycans and lipids in cancer， atherosclerosis， chemical carcinogenesis-reactive oxygen species （ROS） and other related pathways. Conclusion （E）-4-（3，4-dihydroxyphenyl）but-3-en-2-one is the most effective chemical substance in the treatment of pneumonia among the ingredients of Sanghuang， and its mechanism is mainly related to the CYP450 family.

Objective To discuss the differential effects of apolipoprotein E （APOE） gene polymorphism in the neurotoxicity-reactive astrocytes， and to provide the theoretical basis for the study of the pathogenesis of Alzheimer’s disease （AD）. Methods The primary cortical astrocytes from the APOE-knockout mice （APOE ^-/- ） were isolated and cultured in vitro，and the purity of the cells was identified by immunofluorescence staining. The human APOE3 and APOE4 recombinant over-expression plasmids were constructed and separately transfected into the primary APOE ^-/- astrocytes， and the APOE ^-/- primary cells were regarded as control. Western blotting method was used to detect the expression levels of APOE and glial fibrillary acidic protein （GFAP） proteins in the cells； enzyme-linked immunosorbent assay （ELISA） method was used to detect the APOE level in the cellular culture supernatant. The inflammatory models were prepared with the primary astrocytes transfected with APOE3 and APOE4 and co-stimulated with interleukin-1α（IL-1α）， tumor necrosis factor （TNF）， and complement C1q.The cells were divided into APOE3+PBS group， APOE4+PBS group， APOE3+IL-1α+TNF+C1q group， and APOE4+IL-1α+TNF+C1q group. Cell immunofluorescence staining method was used to observe the morphology of the cells in various groups； real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of glypican 4 （Gpc4）， glypican 6 （Gpc6）， thrombospondin 1 （Thbs1）， thrombospondin 2 （Thbs2）， SPARC-like protein 1 （Sparcl1） and glial cell line derived neurotrophic factor （GDNF）， C3，and S100 calcium binding protein B （S100B） mRNA in the cells in various groups； microsphere phagocytosis assay was used to detect the phagocytic capacities of the cells in various groups； Western blotting was used to detect the protein expression levels of B-cell lymphoma 2 （Bcl-2），and cysteinyl aspartate specific protease-3 （Caspase-3） proteins in the cells in various groups. Results Compared with APOE ^-/- group， the expression levels of APOE and GFAP proteins in the cells and the APOE level in the cellular culture supernatant in transfected APOE3 and transfected APOE4 groups were increased （P<0.01）. The fluorescence microscope observation results showed that compared with APOE3+PBS and APOE4+PBS groups， the astrocytic processes in APOE3+IL-1α+TNF+Cq1 group and APOE4+IL-1α+TNF+Cq1 group became shorter and the cell bodies became larger； compared with APOE3+IL-1α+TNF+Cq1 group， the astrocytic processes in APOE4+IL-1α+TNF+Cq1 group were even shorter. Compared with APOE3+PBS and APOE4+PBS groups， the expression levels of Gpc4， Gpc6， Thbs1， Thbs2，and Sparcl1 mRNA in the cells in APOE3+IL-1α+TNF+Cq1 group and APOE4+IL-1α+TNF+Cq1 group were significantly decreased （P<0.01）； compared with APOE3+IL-1α+TNF+Cq1 group， the expression levels of Gpc4， Gpc6，Thbs1， Thbs2， and Sparcl1 mRNA in the cells in APOE4+IL-1α+TNF+Cq1 group were significantly decreased （P<0.05 or P<0.01）. Compared with APOE3+PBS and APOE4+PBS groups， the expression levels of GDNF mRNA in the cells in APOE3+IL-1α+TNF+Cq1 group and APOE4+IL-1α+TNF+Cq1 group were decreased（P<0.01），and the expression levels of C3 and S100B mRNA were increased（P<0.01）；compared with APOE3+IL-1α+TNF+Cq1 group， the expression level of GDNF mRNA in the cells in APOE4+IL-1α+TNF+Cq1 group was decreased（P<0.05），and the expression levels of C3 and S100B mRNA were increased（P<0.05）. Compared with APOE3+PBS group and APOE4+PBS group，the numbers of hagocytosis of microspheres in the cells in APOE3+IL-1α+TNF+Cq1 group and APOE4+IL-1α+TNF+Cq1 group were significantly decreased； compared with APOE3+IL-1α+TNF+Cq1 group，the number of hagocytosis of microspheres in the cells in APOE4+IL-1α+TNF+Cq1 group was significantly decreased. Compared with APOE3+PBS group and APOE4+PBS group，the expression levels of Bcl-2 protein in the cells in APOE3+IL-1α+TNF+Cq1 group and APOE4+IL-1α+TNF+Cq1 group were decreased （P<0.05 or P<0.01）and the expression levels of Caspase-3 protein were significantly increased（P<0.01）；compared with APOE3+IL-1α+TNF+Cq1 group， the expression level of Bcl-2 protein in the cells in APOE4+IL-1α+TNF+Cq1 group was decreased（P<0.01），and the expression level of Caspase-3 protein was increased（P<0.05）. Conclusion The APOE4 genotype has a stronger ability to induce the inflammatory factors compared with APOE3；it can lead to a neurotoxicity-reactive astrocyte phenotype，increase the neurotoxicity，affect the astrocyte apoptosis， and aggravate the neuron damage.

Objective To discuss the expression of programmed cell death-ligand 1 （PD-L1） in the oral squamous cell carcinoma （OSCC） cells and its effect on biological behavior of the OSCC CAL27 cells， and to clarify the possible mechanism. Methods Western blotting method was used to detect the expression levels of PD-L1 protein in the oral epithelial HOK cells and OSCC CAL27， TCA8113， and SCC15 cells； immunofluorescence staining method was used to detect the expression and localization of PD-L1 protein in the CAL27 cells. The CAL27 cells were divided into control group （transfected with si-NC） and si-PD-L1 group （transfected with si-PD-L1）. Western blotting method was used to detect the interference efficiency of the cells in two groups； CCK-8 assay was used to detect the proliferative activities of the cells in two groups at different time points； plate clone formation assay was used to detect the numbers of clone formation of the cells in two groups； cell scratch healing assay was used to detect the scratch healing rates of the cells in two groups； Transwell chamber assay was used to detect the numbers of migration and invasion cells in two groups. Results The expression level of PD-L1 protein in the OSCC cells was higher than that in the HOK cells （P<0.05 or P<0.01）； PD-L1 expressed in the cytoplasm and nucleus of the CAL27 cells. The CCK-8 assay and plate clone formation assay results showed that compared with control group， the proliferative activities of the CAL27 cells in si-PD-L1 group at different time points were significantly decreased（P<0.05 or P<0.01）， and the numbers of clone formation were significantly decreased （P<0.01）. The cell scratch healing assay results showed that compared with control group， the scratch healing rates of the cells in si-PD-L1 group were significantly decreased （P<0.05 or P<0.01）. The Transwell chamber assay results showed that compared with control group， the numbers of migration and invasion cells in si-PD-L1 group were significantly decreased （P<0.01）. Conclusion The expression of PD-L1 in the OSCC cells is higher than that in normal oral epithelial cells， and knocking down PD-L1 expression can inhibit the proliferation， clone formation， migration and invasion capabilities of the OSCC cells.

Objective To disuss the effect of glucagon like peptide-1 receptor （GLP-1R） agonist Exendin-4（E4） on injury of cardiomyocytes in the hyperglycemia and hyperlipidemia（HGHL） environment， and to clarify its related mechanism. Methods The H9C2 cardiomyocytes of the rats were divided into control group， HGHL group， HGHL+E4 group， HGHL+E4+iron death agonist（Erastin） group.The HGHL model of the H9C2 cells was induced by 33 mmol·L^－1 glucose and 500 μmol·L^－1 palmitate，and the cells in HGHL+E4 group and HGHL+E4+Erastin group were treated with 100 nmol·L^－1 E4 and 5 μmol·L^－1 Erastin. CCK-8 assay was used to detect the survival rates of cells in various groups； Hoechst 33258 fluorescence staining was used to observe the apoptosis of cells in various groups； flow cytometry was used to detected the apoptotic rates of cells in various groups；2'，7'-dichlorodihydrofluorescein diacetate（DCFH-DA） fluorescence probe was used to detect the levels of reactive oxygen species （ROS） in the cells in various groups； the corresponding kit was used to detect the levels of glutathione （GSH） and malondialdehyde （MDA） in cells in various groups；JC-1 staining was used to detect the mitochondrial membrane potential （MMP） in cells in various groups； FerroOrange fluorescence probe was used to detect the levels of Fe²⁺ in the cells in various groups； Western blotting method was used to detect the expression levels of glutathione peroxidase 4 （GPX4） and solute carrier family 7 member 11（SLC7A11） in the cells in various groups. Results Compared with control group， the survival rate of cells in HGHL group was decreased （P<0.05），and the cells were fragmented and collapsed， which showed apoptotic morphology；the apoptotic rate and the levels of ROS and MDA in the cells were increased（P<0.05）， the levels of GSH and MMP were decreased（P<0.05）， the level of Fe²⁺ was increased（P<0.05），and the expression levels of GPX4 and SLC7A11 proteins were decreased（P<0.05）.Compared with HGHL group， the survival rate of the cells in HGHL+E4 group was increased （P<0.05），and the apoptosis was decreased；the apoptotic rate and the levels of ROS and MDA were decreased（P<0.05）， while the levels of GSH and MMP were increased（P<0.05）， the level of Fe²⁺ was decreased（P<0.05）， and the expression levels of GPX4 and SLC7A11 proteins were decreased （P<0.05）.Compared with HGHL+E4 group， the survival rate of the cells in HGHL+E4+Erastin group was decreased（P<0.05）， the cell fragmentation and wrinkling were obviously decreased，the apoptotic rate and the levels of ROS and MDA were increased（P<0.05）， the levels of GSH and MMP were decreased（P<0.05）， the level of Fe²⁺ was increased（P<0.05），and the expression levels of GPX4 and SLC7A11 proteins were decreased（P<0.05）. Conclusion GLP-1R agonists can reduce the cardiomyocyte injury in the HGHL environment， and the protective effect may be related to inhibiting the iron death.

Objective To construct the sex-determined region Y-box 17 （SOX17） over-expression lentiviral vector， and to construct the cell line stably over-expressing SOX17 by using the PC12 cells infected with SOX17 over-expression lentivirus. Methods The over-expression sequence of SOX17 was designed and synthesized based on the National Center for Biotechnology Information （NCBI） Database， and was connected to the GV492 lentiviral vector after being doubly digested with BamHⅠ and AgeⅠ enzymes to construct the GV492-SOX17 over-expression recombinant plasmid. The agarose gel electrophoresis was used to identify the PCR products， and the positive bacteria carrying the GV492-SOX17 over-expression recombinant plasmid were selected， cloned and sequenced. The GV492 empty plasmid and GV492-SOX17 over-expression recombinant plasmid were transfected into the human embryonic kidney HEK 293T cells， respectively. After transfected for 48 h， the GV492 control lentivirus and GV492-SOX17 over-expression lentivirus were packaged and the viral titer was detected. The PC12 cells were divided into blank group， GV492 control group， and GV492-SOX17 group. The cells in blank group was not treated， and the cells in GV492 control and GV492-SOX17 groups were infected with the corresponding lentivirus （multiplicity of infection = 100）， and the PC12 cells successfully infected with lentivirus were selected with 10 mg·L^－1 puromycin. The growth state and green fluorescence expression of the PC12 cells were observed under fluorescence microscope；real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression level of SOX17 mRNA in the PC12 cells in various groups；Western blotting method was used to detect the expression level of SOX17 protein in the PC12 cells in various groups. Results The gene fragment length of GV492-SOX17 over-expression recombinant plasmid was about 744 bp， and the sequence of GV492-SOX17 over-expression recombinant plasmid gene was identical to the designed and synthesized SOX17 over-expression sequence. The titers of GV492 control lentivirus and GV492-SOX17 over-expression lentivirus were both 2.5×108 TU·mL^－1. The growth state of the PC12 cells in various groups was good and the green fluorescence was expressed. The RT-qPCR results showed that the expression level of SOX17 mRNA in the cells in GV492-SOX17 group was significantly higher than those in blank group and GV492 control group（P<0.01）. The Western blotting results showed that there was a specific band appeared at a relative molecular mass of 44 000， suggesting the SOX17 protein was successfully expressed in the PC12 cells； compared with blank group and GV492 control group， the expression level of SOX17 protein in the cells in GV492-SOX17 group was increased （P<0.05）. Conclusion The GV492-SOX17 lentiviral expression vector is successfully constructed and the PC12 cell line stably over-expressing GV492-SOX17 is established.

Objective To analyze the improvement effect of monk fruit on diabetic nephropathy（DN） by network pharmacology，and to elucidate its possible related mechanism. Methods The Traditional Chinese Medicine Systems Pharmacology（TCMSP） Database was used to detect the active ingredients and their targets of monk fruit； the DN target genes were screened out by DisGeNET Database and Genecards Database； the key targets of monk fruit against DN were obtained by comparing the monk fruit with DN targets； protein-protein interaction （PPI） network diagram was constructed by STRING Database and Cytoscape software； Gene Ontology （GO） functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathway enrichment analysis were performed by Cytoscape software.Molecular docking technology was used to predict the binding abilities of the core targets and the main active ingredients of monk fruit. Results The TCMSP Database combined with the selection criteria was used to screen out a total of five active ingredients of monk fruit （ZINC03860434， Perlolyrine， beta-sitosterol， Kaempferol， and Flazin） as well as 85 targets represented by serine/threonine protein kinase 1 （AKT1），transcription factor RELA， c-Jun N-terminal kinase （JUN），and tumor necrosis factor （TNF）. Among them， Kaempferol contained the most targets.Among the 85 targets， 34 were associated with DN.The GO functional enrichment analysis mainly included biological process（BP） such as oxidative stress， regulation of inflammation and apoptosis， and cell signaling transduction.The KEGG enrichment analysis included advanced glycosylation end product（AGE）-receptor of AGE （AGE-RAGE） signaling pathway， TNF signaling pathway， and C-type lectin receptor signaling pathway.The results molecular docking technology of the main active ingredients of monk fruit and DN target proteins showed that 5 kinds of molecular docking engergy were －8.00－－5.00 kJ·mol^－1. Conclusion Kaempferol is the most effective active ingredient in the monk fruit for the treatment of DN， and its mechanism is mainly related to anti-inflammatory.

Objective To discuss the effect of Sirtuins（Sirt） protein family on the male reproductive system damage cell and animal models induced by bisphenol A （BPA）， and to clarify its effect on the male reproductive system damage induced by BPA. Methods Forty Kunming mice were randomly divided into control group， low dose of BPA group （given 3 mg·kg^－1·d^－1 BPA）， medium dose of BPA group （given 30 mg·kg^－1·d^－1 BPA）， and high dose of BPA group （given 300 mg · kg^－1·d^－1 BPA），and there were 10 mice in each group. The mice in low， medium， and high doses of BPA groups were gavaged with corn oil （0.01 mL·g^－1） mixed with corresponding concentrations of BPA， while the mice in control group were given 0.01 mL·g^－1 corn oil. After 5 weeks， the body weights， testis indexes， and epididymal indexes of the mice in various groups were detected； the sperm qualities of the mice in various groups were detected by using computer assisted semen analysis （CASA） system； the morphology of testis tissue of the mice in various groups was observed by HE staining； the expression levels of Sirt1－Sirt7 proteins in testis tissue of the mice in various groups were detected by Western blotting method. The GC-2 cells were divided into 0， 20， 40， and 80 μmol·L^－1 BPA groups （treated with 0， 20， 40，and 80 μmol·L^－1 BPA）. The proliferation rates of the GC-2 cells in various groups were detected by EdU staining； the percentages of the GC-2 cells at different cell cycles and the apoptotic rates of GC-2 cells in various groups were detected by flow cytometry；the expression levels of Sirt1－Sirt7 proteins in the GC-2 cells in various groups were detected by Western blotting method. Results Compared with control group， the testis index of the mice in high dose of BPA group was decreased （P<0.05）； compared with control group， the percentage of immobile sperm of the mice in high dose of BPA group was increased （P<0.01）， the percentage of rapid progressive sperm was decreased （P<0.01）， the percentage of medium progressive sperm was decreased （P<0.05）， and the deformity rate of the sperm was increased （P<0.01）. The HE staining results showed that the number of spermatogenic cells at all levels in the seminiferous tubules of the mice in different doses of BPA groups showed a decreasing and loosely arranged trend. Compared with control group， the expression level of Sirt6 protein in testis tissue of the mice in low dose of BPA group was decreased （P<0.01）， while the expression levels of Sirt1， Sirt2， and Sirt6 proteins in testis tissue of the mice in medium dose of BPA group mice were decreased （P<0.01），the expression levels of Sirt1， Sirt2， Sirt3， Sirt4， Sirt5， Sirt6， and Sirt7 proteins in testis tissue of the mice in high dose of BPA group were decreased （P<0.05 or P<0.01）. The EdU staining results showed that compared with 0 μ mol·L^－1 BPA group， the proliferation rates of the GC-2 cells in 20， 40， and 80 μmol·L^－1 BPA groups were decreased （P<0.01）. The flow cytometry results showed that after treated for 48 h， compared with 0 μmol·L^－1 BPA group， the apoptotic rates of the GC-2 cells in 20， 40， and 80 μmol·L^－1 BPA groups were increased （P<0.01）. The Western blotting assay results showed that after treated for 24 h， compared with 0 μmol·L^－1 BPA group， the expression levels of Sirt4 and Sirt7 proteins in GC-2 cells of the mice in 20 μmol·L^－1 BPA group were decreased （P<0.05 or P<0.01）， while the expression levels of Sirt4 and Sirt7 proteins in GC-2 cells of the mice in 80 μmol·L^－1 BPA group decreased （P<0.05 or P<0.01）. After treated for 48 h， compared with 0 μmol·L^－1 BPA group， the expression levels of Sirt2， Sirt3， Sirt4， and Sirt6 proteins in GC-2 cells of the mice in 20 μmol·L^－1 BPA group were decreased （P<0.05 or P<0.01）， while the expression levels of Sirt1， Sirt2， Sirt3， Sirt4， and Sirt6 proteins in GC-2 cells of the mice in 40 μmol·L^－1 BPA group were significantly decreased （P<0.05 or P<0.01），and the expression levels of Sirt 5 and Sirt 6 proteins were decreased （P<0.05 or P<0.01）. Conclusion The expression levels of Sirt1－Sirt7 proteins in the male reproductive injury cells and animal models induced by BPA are decreased， which exacerbates the male reproductive system damage induced by BPA.

Objective To discuss the effect of shear wave elastography （SWE） on the differential diagnosis of benign and malignant breast non-mass lesion （NML）， and to clarify its clinical significance. Methods A total of 158 patients with breast NML were selected and divided into benign lesion group （ n=83） and malignant lesion group（ n=75） according to the postoperative pathological type of the patients.The routine ultrasound and SWE examinations were conducted before operation， and the SWE parameters of the lesions， including the maximum （E_max）， average （E_mean），minimum （E_min）， standard deviation （E_sd） and whether the stiff rim sign presented or not at 1， 2 and 3 mm around the lesion were recorded. The SWE characteristics and differences in SWE parameters of the NML patients between two groups were analyzed， and the receiver operating characteristic curve（ROC） was plotted， and the area under curve （AUC）， specificity and sensitivity were calculated. Results There were statistically significant differences in age， lesion size， palpation， and whether menopause had crested of the NML patients between two groups（ P<0.01）. All the lesions of the patients in two groups appeared as hyperechoic and irregular in shape. Compared with benign lesion group， the percentage of the patients with non-parallel， calcification and posterior echo attenuation in malignant lesion group was significantly increased （ P<0.01）， and the percentage of the patients without blood supply was significantly decreased （ P<0.01）. Compared with benign lesion group， the E_max， E_mean， and E_sd of the NML patients in malignant lesion group were significantly increased （ P<0.01）， and the E_max， E_mean， and E_sd at 1， 2， and 3 mm around the lesion were all increased （ P<0.01）. The stiff rim sign manifested as a circular or semi-circular red region around the lesions of the patients in malignant lesion group. Compared with benign lesion group， the percentage of hard rim sign of the NML patients in malignant lesion group was significantly increased （ P<0.01）.Compared with inside the lesion，the E_min at 1，2 and 3 mm around the lesion of the NML patients in benign lesion group and malignant lesion group were decreased（ P<0.01）. The AUC of E_max at 1， 2 and 3 mm around the lesion were 0.872， 0.860， 0.873 and 0.866，respectively； the AUC of E_mean were 0.796， 0.822， 0.820 and 0.815，respectively； the AUC of E_sd were 0.832， 0.857， 0.859， and 0.842，respectively；the sensitivity and specificity of E_max inside the lesion were 85.33% and 83.13%，respectively；the sensitivity and specificity of E_max at 1 mm around the lesion were 82.67% and 80.72%，respectively； the sensitivity and specificity at 2 mm around the lesion were 78.67% and 87.95%，respectively； the sensitivity and specificity at 3 mm around the lesion were 80.00% and 86.75%，respectively； the specificity of E_sd at 2 mm around the lesion was highest （93.33%）. Conclusion The SWE parameters inside and around the lesion and the manifestation of stiff rim sign can provide the reference for the differential diagnosis of benign and malignant breast NML， and have good diagnostic performance.

Objective To screen out the key genes affecting lung adenocarcinoma （LUAD） through bioinformatics methods，and to analyze their biological functions and the influences on the LUAD prognosis. Methods The GSE118370 and GSE136043 chip data were downloaded from the Gene Expression Omnibus（GEO） Database. The LUAD-related data were selected from the The Cancer Genome Atlas（TCGA） Database. R software was used to analyze the co-expressed differentially expressed genes （DEGs）； clusterProfile R package was utilized for Gene Ontology （GO） functional enrichment analysis； DAVID Database was used for the Kyoto Gene and Genome Encyclopedia （KEGG） signaling pathway enrichment analysis； STRING Database was used to construct the protein-protein interaction （PPI） network； Cytoscape was used to screen out the top 10 key genes； GEPIA Database and Human Protein Atlas（HPA） Database were used to analyze the expressions of key genes mRNA and protein in normal lung tissue and LUAD tissue， and their expressions in LUAD tissues with different stages；immune infiltration analysis and survival analysis were used to analyze the correlation between the expressions of key genes and the survival time of the patients. Results In total， 428 DEGs were screened out. The GO functional analysis results showed that the DEGs of LUAD were mainly enriched in biological process（BP） such as epithelial-mesenchymal transition （EMT）， cellular component（CC） such as the cell base， and molecular function（MF） such as extracellular matrix （ECM） structure formation.The KEGG signaling pathway analysis results showed that the DEGs of LUAD were mainly enriched in the pathways like cytokine receptor interactions.topoisomerase Ⅱ alpha（TOP2A），abnormal spindle microtubule assembly（ASPM），cyclin B1，（CCNB1），cell division cycle associated 8（CDCA8），baculoviral IAP repeat containing 5（BIRC5），aurora A（AURKA），kinesin family member 20A（KIF20A），centrosomal protein 55（CEP55），centromere protein F（CENPF），and targeting protein for xklp2（TPX2） were selected as the key genes. Compared with normal lung tissue， the expression levels of TOP2A， CCNB1， CDCA8， BIRC5， AURKA， KIF20A， CEP55， CENPF， and TPX2 mRNA in lung tissue of the LUAD patients were increased （P<0.01）， and the expression levels of TOP2A， CCNB1， CDCA8， BIRC5， AURKA， KIF20A， CEP55， CENPF， and TPX2 proteins were increased. There were significant differences in the expression levels of CCNB1， CDCA8， BIRC5， AURKA， KIF20A， CEP55， and TPX2 mRNA in the LUAD tissue with different stages （P<0.01）. Compared with the LUAD patients with stage Ⅰ，stage Ⅱ，and stage Ⅲ，the expression levels of CCNB1， CDCA8， AURKA， KIF20A， CEP55， and TPX2 mRNA in lung tissue of the LUAD patients with stage Ⅳ were increased（P<0.01）； compared with the LUAD patients with stage Ⅰ，stage Ⅱ， and stage Ⅳ，the expression level of BIRC5 mRNA in lung tissue of the LUAD patients with stage Ⅲ was increased （P<0.01）. The expressions of 10 key genes were negatively correlated with the B-lymphocyte infiltration （－0.253≤r≤－0.014， P<0.01）； the expressions of TOP2A， ASPM， CDCA8， BIRC5， CEP55， CENPF， and TPX2 were positively correlated with the neutrophil infiltration （0.049≤r≤0.165，P<0.01）； the expressions of CCNB1 and AURKA were negatively correlated with the CD4 T lymphocyte， macrophage， and dendritic cell infiltration （－0.210≤r≤－0.100，P<0.01）. The high expression of CDCA8 increased the risk of LUAD deterioration （P<0.01）， and the high expressions of TOP2A， CCNB1， CDCA8， BIRC5， AURKA， KIF20A， CEP55， CENPF， and TPX2 increased the death risk of the patients（P<0.01）. Conclusion TOP2A， ASPM， CCNB1， CDCA8， BIRC5， AURKA， KIF20A， CEP55， CENPF， and TPX2 are the key genes involved in the development and progression of LUAD. They may promote the development of LUAD by accelerating the EMT process， and their high expressions suggest the poor prognosis and elevate the death risk of the LUAD patients.

Objective To discuss the effect of Jiegeng Yuanshen Tang（JGYST） on airway tissue inflammation and mucus secretion in the mice with allergic asthma， and to clarify the related mechanism. Methods Forty male C57BL/J mice were randomly divided into control group， JGYST group， ovalbumin （OVA） group， and OVA + JGYST group. The mice in OVA group and OVA +JGYST group were sensitized with 50 μg OVA via intraperitoneal injection twice weekly， followed by 20 μg OVA nasal drops daily for 7 d to induce asthma；the mice in OVA +JGYST group were gavaged with 200 μL JGYST 1 h before each OVA challenge， and the administration lasted for 7 d； the mice in control group were given equivalent dose of PBS via intraperitoneal injection， nasal drops， and gavage； the mice in JGYST group were given the same dose of PBS for intraperitoneal and nasal administration and gavaged with the same dose of JGYST. The pathomorphology of lung tissue of the mice in various groups was observed by HE staining and periodic acid-Schiff （PAS） staining， and the inflammation and PAS scores were calculated； flow cytometry method was used to detect the numbers of eosinophils， neutrophils， helper T lymphocyte 1 （Th1） cells， helper T lymphocyte 2 （Th2） cells， and dendritic cells （DCs）， as well as the percentage of mature DCs and level of reactive oxygen species （ROS） in lung tissue of the mice in various groups；real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of interleukin-4 （IL-4）， interleukin-10 （IL-10）， and tumor necrosis factor-α （TNF-α） mRNA in lung tissue of the mice in various groups. Results The HE and PAS staining results showed that the mice in control group had intact airway and alveolar structure，without infiltration of inflammatory cells or mucus secretion； compared with control group， there was a large number of infiltrating inflammatory cells in airway tissue of the mice in OVA group，and the inflammation and PAS scores were increased （P<0.01）； compared with OVA group， the infiltration of inflammatory cells in airway tissue of the mice in JGYST group and OVA + JGYST group was decreased， and the inflammation and PAS scores were significantly decreased （P<0.01）. The flow cytometry results showed that compared with control group， the numbers of eosinophils， Th2 cells， and DCs in lung tissue of the mice in OVA group were increased （P<0.05 or P<0.01）， and the percentage of mature DCs and level of ROS were significantly increased （P<0.01）； compared with OVA group， the numbers of eosinophils， Th2 cells， and DCs in lung tissue of the mice in JGYST group and OVA + JGYST group were decreased （P<0.01）， and the percentage of mature DCs and level of ROS were significantly decreased （P<0.01）. The RT-qPCR results showed that compared with control group， the expression levels of IL-4， IL-10， and TNF-α mRNA in lung tissue of the mice in OVA group were increased （P<0.01）； compared with OVA group， the expression levels of IL-4 and TNF-α mRNA in lung tissue of the mice in JGYST group and OVA + JGYST group were decreased （P<0.01）， while the expression level of IL-10 mRNA was increased （P<0.01）. Conclusion JGYST can alleviate the airway tissue inflammation and mucus secretion in the mice with allergic asthma，and its mechanism may be related to reducing the number of Th2 cells and DCs， decreasing the ROS level and expression level of proinflammatory cytokine， and increasing the expression level of anti-inflammatory cytokine.

Objective To screen the aging genes closely associated with pelvic organ prolapse （POP） by bioinformatics techniques， and to clarify the potential clinical significance and value of key genes. Methods Gene Expression Omnibus （GEO） Database was used to download the datasets GSE53868 and GSE151188 for POP-related genes with the keyword “pelvic organ prolapse”. The aging-related genes were obtained from Aging Atlas， CellAge， and the Human Ageing Genomic Resources （HAGR） Databases；the intersection of genes related with POP in two groups provided a list of differentially expressed genes （DEGs） associated with aging in POP； gene Set Enrichment Analysis （GSEA） was conducted with R software version 4.2.1； Gene Ontology （GO） functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathway enrichment analysis of DEGs were conducted by the Database for Annotation， Visualization and Integrated Discovery （DAVID）； the protein-protein interaction （PPI） network was constructed with Cytoscape 3.9.1 software；the top 10 Hub genes were selected by cytoHubba plugin； the infiltration of 22 types of immune cells in the patients in POP group and control group was analyzed by CIBERSORT deconvolution method using R software；the key genes were further screened by LASSO regression algorithm； the correlation and diagnostic efficacy between key genes and immune cell infiltration were analyzed. Results From the Aging Atlas， CellAge， and HAGR Databases， 724 aging-related genes were identified. Intersection with the POP expression profile yielded an aging gene expression matrix related to POP containing 624 genes， and 29 POP-related DEGs were identified after differential analysis， including 2 upregulated genes and 27 downregulated genes. The GSEA results showed that the upregulated pathways were mainly related to diabetes and cellular senescence， whereas the downregulated pathways included Alzheimer’s disease and hypoxia-inducible factor-1 （HIF-1） signaling pathways.The GO functional enrichment analysis mainly enriched in the biological processes such as the response of the cells to lipopolysaccharide， inflammatory response， and negative regulation of cell proliferation. The KEGG signaling pathway enrichment analysis mainly enriched in interleukin-17 （IL-17）， tumor necrosis factor （TNF）， and nuclear factor-kappa B （NF-κB） signaling pathways. The PPI network analysis got 10 Hub genes including interleukin-6 （IL-6）， interleukin-1B （IL-1B）， prostaglandin-endoperoxide synthase 2 （PTGS2）， and NF-kappa-B inhibitor alpha （NFKBIA）. The CIBERSORT deconvolution method results showed a relatively higher infiltration proportion of neutrophils and activated mast cells in the patients in POP group， the activated mast cells had a positive correlation with most of the DEGs （r>0.5） and the macrophages had a significant positive correlation with IL-1B （r>0.6）. The key genes Jun D proto-oncogene （JUND）， Snail homolog 1 （SNAI1）， amphiregulin （AREG）， Lamin A/C （LMNA）， and superoxide dismutase 2 （SOD2） selected by LASSO regression analysis had high diagnostic efficacies， and the area under receiver operating characteristic curve （ROC） （AUC） were all greater than 0.75. Conclusion During the aging process，the genes such as JUND，SNAI1，AREG，LMNA，and SOD2 may participate in the pathophysiology of POP through various pathways，including inflammation-related pathways，transcription regulation，and affecting collagen secretion and metabolism，thereby influence the connective tissue support function and promote the occurrence and development of POP.

Objective To identify the key genes associated with the early diagnosis and poor prognosis of hepatitis B virus-associated hepatocellular carcinoma （HBV-HCC） by using the bioinformatics methods， and to elucidate the underlying molecular mechanism of occurence and development of HBV-HCC. Methods Gene Expression Omnibus （GEO） Database was used to retrieval “hepatitis B induced HCC”； the Gene Dataset GSE121248 was downloaded， the differentially expressed genes （DEGs） were screened by the “limma” Data Package in R Software， and the DEGs were enriched by using the “clusterProfiler” Data Package for Gene Ontology （GO） functional analysis and the Kyoto Genes and Genome Encyclopedia （KEGG） signaling pathway enrichment analysis； STRING Database and Cytoscape Software were used to establish the protein-protein interaction（PPI） network and the key genes were screened out.Gene Expression Profiling Interactive Analysis（GEPIA）， Kaplan Meier-Plotter，and Human Protein Atlas（HPA）Databases were used to verify the key genes and the expression levels of proteins；the infiltration of the immune cells was analyzed based on the “CIBERSORT” Data Package. Results A total of 574 DEGs were identified，including 173 up-regulated genes and 401 down-regulated genes. The GO functional enrichment analysis results showed that DEGs were mainly enriched in the biological processes such as small molecule metabolism， signal transduction， immune response， inflammatory response，and so on； the KEGG signaling pathway enrichment analysis results showed that the DEGs were mainly enriched in retinol metabolism， cytochrome P450 metabolic pathway of exogenous drugs， and chemical carcinogenesis，and so on. The PPI network results showed that cell division cycle 20（CDC20），cyclin dependent kinase 1（CDK1）， cyclin A2（CCNA2）， pindle checkpoint protein（BUB1B）， topoisomerase Ⅱ α（TOP2A）， discs large homolog associated related protein 5（DLGAP5）， abnormal spindle-like microcephaly associated protein（ASPM）， centrosomal protein 55（CEP55）， kinesin superfamily 11（KIF11），and kinesin superfamily 20A（KIF20A） were the key genes. The GEPIA Database analysis results showed that these above 10 key genes were highly expressed in the HCC patients. The Kaplan Meier survival curve showed that the overall survivals of the HCC patients with high expression of key genes were shorter than those of the HCC patients with low expression of key genes. Conclusion The genes related to cell cycle and viral oncogenesis （CDC20， CDK1， CCNA2， and BUB1B） are closely associated with the occurence and development and poor prognosis of the HBV-HCC patients， which may become the diagnostic markers and new targets for the treatment.

Objective To discuss the effect of ganoderic acid A （GAA） on the proliferation， apoptosis， and migration ability of the PC-9 cells， and to clarify its mechanism. Methods The non-small cell lung cancer （NSCLC） PC-9 cells were cultured in vitro and divided into control group （without GAA）， low dose of GAA group （0.1 mmol·L^－1 GAA）， and high dose of GAA group （0.5 mmol·L^－1 GAA）. MTT method was used to detect the proliferation rates of the PC-9 cells in various groups； flow cytometry was used to detect the apoptotic rates of the PC-9 cells in various groups； scratch wound healing assay was used to detect the scratch healing rates of the PC-9 cells in various groups； Transwell chamber assay was used to detect the migration abilities of PC-9 cells in various groups； the expression levels of vascular endothelial growth factor （VEGF） and hypoxia-inducible factor-1α （HIF-1α） mRNA and protein in the PC9 cells in various groups were detected by real-time fluorescence quantitative PCR （RT-qPCR） and Western blotting methods. Results The MTT assay results showed that compared with control group， the proliferation rate of the cells in low dose of GAA group was decreased after treated for 48 and 72 h （P<0.05）； after treated for 24， 48， and 72 h， the proliferation rate of the cells in high dose of GAA group was decreased （P<0.05）； compared with low dose of GAA group， the proliferation rate of the cells in high dose of GAA group was decreased after treated for 24， 48， and 72 h （P<0.05）.The flow cytometry results showed that compared with control group and low dose of GAA group， the apoptotic rate of the cells in high dose of GAA group was increased （P<0.05）. The cell scratch wound healing assay results showed that compared with control group and low dose of GAA group， the scratch wound healing rate of the cells in high dose of GAA group was decreased （P<0.05）. The Transwell chamber assay results showed that compared with control group and low dose of GAA group， the number of migration cells in high dose of GAA group was decreased （P<0.05）. The RT-qPCR and Western blotting results showed that compared with control group and low dose of GAA group， the expression levels of HIF-1α and VEGF mRNA and proteins in the cells in high dose of GAA group were decreased （P<0.05）. Conclusion High dose of GAA can inhibit the proliferation of the PC-9 cells and promote the apoptosis， and its mechanism may be related to regulating the expressions of VEGF and HIF-1α mRNA and proteins.

objective To prepare a composite photocrosslinked hydrogel containing zeolite imidazole framework-8 （ZIF-8），and to evaluate its in vitro cytotoxicity， drug release capability， and antimicrobial propertie. Methods The ZIF-8 particles were synthesized by hydrothermal method， and the microstructure characteristic was observed under scanning electron microscope （SEM）. The particles were mixed with the gelatin methacryloyl （GelMA） with the mass fraction of 0.2% to obtain the composite hydrogel GelMA-Z. The atomic absorption spectroscope was used to detect the cumulative zinc ion （Zn²⁺） release amounts in GelMA-Z at different time points.The NIH-3T3 cells were co-cultured with GelMA-Z for 1， 3， and 7 d；the viabilities of the cells in various groups were detected by CCK-8 assay； the GelMA-Z was co-cultured with Escherichia coli （E. coli） and Staphylococcus aureus （S. aureus） for 6，12，and 24 h and divided into control group， GelMA group， and GelMA-Z group.The bacterial activities of the cells in various groups at different time points were detected by microplate reader； the bacterial formation and the presence of live/dead becterial staining condition were detected by plate antibacterial experiment and live/dead bacterial staining method. Results The SEM observation results showed that the hydrothermally synthesized ZIF-8 particles had the uniform particle sizes.The atomic absorption spectroscope results showed that Zn²⁺ in GelMA-Z showed an initial burst phase within 1 d， followed by a slow release，and reached the equilibrium around 7 d.Compared with control group，the viabilities the cells in GelMA group and GelMA-Z group were above 90% on the 1st， 3rd， and 7th days， but there was no significant difference（P>0.05）.The bacterial activity detection results showed that when co-cultured with bacteria for 6，12，and 24 h， compared with control group and GelMA group，the bacterial activities of the E. coli and S. aureus in GelMA-Z group were decreased （P<0.05）. The plate antibacterial experiment results showed that the number of bacterial formation in GelMA-Z group was fewer than those in control group and GelMA group. The live/dead bacterial staining results showed that in GelMA-Z group，there was a large number of red fluorescence stained dead bacteria； in control group and GelMA group， there was a large number of green fluorescence stained live bacteria. Conclusion The GelMA hydrogel loaded with ZIF-8 particles can achieve the in situ photocrosslinking and possesses good Zn²⁺ release capability and antimicrobial activity， and it is a novel hydrogel dressing for treatment of the infected wounds.

Objective To discuss the method of primary culture and activity evaluation system of the neural stem cells （NSCs） of the fetal rats， and to confirm the optimal subculture conditions of the NSCs. Methods The SD rats with 11－14 d gestation were chosen，and the primary NSCs of the fetal rats were extracted. Nestin immunofluorescence staining was used to identify the NSCs. The NSCs were subcultured at the cell densities of 1.0×10⁴，2.0×10⁴，6.0×10⁴，1.0×10⁵， 1.6×10⁵ and 2.0×10⁵ mL^－1， then the morpholgy of the neurospheres was observed 48 h after passage and the diameter of the neurospheres was calculated. The survival rates of NSCs in the neurospheres with different diameters were detected by live-dead cell staining. Results The expression of nestin in the NSCs was positive， indicating that the cultured cells were the NSCs. The NSCs showed aggregation growth and strong cell-cell adhesion pattern forming neurospheres with an average diameter of （152.72±47.52） μm， and there were few single cells scattered between the neurospheres. Compared with 2.0×10⁴ mL^－1 subculture density，the diameters of the neurospheres at the subculture densities of differences 6.0×10⁴ mL^－1 and 1.0×10⁵ mL^－1 were increased （P<0.05）. There were no significant differences in the diameters of the neurospheres at the subculture densities of 1.0×10⁵ mL^－1， 1.6×10⁵ mL^－1， and 2.0×10⁵ mL^－1 （P>0.05）. Compared with neurospheres with diameters of 0—40 μm， 40—60 μm， 60—80 μm，and 80—100 μm， the survival rate of NSCs in the neurospheres with the diameters of 100—200 μm was decreased （P<0.05）. Conclusion The neurospheres with the diameters of 80—100 μm when subcultured at the density of 1.0×10⁵ mL^－1 can effectively improve the efficiency and activity of subculture of the NSCs.

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 The human-derived antimicrobial peptide（AMPs） histatin 5 （Hst5） and LL-37 were used as the parents to design a novel heterozygous peptide H5LL， and to construct a recombinant lactic acid bacteria expression vector by genetic engineering technology to achieve the efficient and safe expression of the AMPs. Methods Bioinformatics method was used to predict the physicochemical parameters， hydrophilicity/hydrophobicity， shear sites， phosphorylation sites， signal peptides and transmembrane regions， subcellular localization， and secondary structure of H5LL； the target sequence and the empty plasmid pMG36e were doubly digested by HindⅢ and KpnⅠ， then the lactic acid bacteria recombinant expression vector pMG36e-H5LL was constructed and cloned to obtain the recombinant plasmid containing the target genes. Results H5LL had a high possibility of being an AMPs， contained 36 amino acids， the relative molecular weight was 4 625 380，the total charge number was +10， which had hydrophilicity； there were 3 phosphorylation sites and there was no glycosylation site； H5LL belonged to intramembrane protein， without transmembrane region and signal peptide；the subcellular localization prediction results showed that the possibility of mitochondrial targeting peptide was 0.333. The α-helix junction and β-turned angle accounted for 52.78% and 22.22% in the secondary structure， and the number of α-helix was the highest in the tertiary structure. The PCR electrophoresis results of recombinant plasmid contained target gene showed a single specific band at 138 bp； the recombinant plasmid double digestion electrophoresis results showed the clear bands at 136 and 3 500 bp. Conclusion The novel heterogeneous peptide H5LL is designed and synthesized with high stability， antibacterial activity and low toxicity；the recombinant lactic acid bacteria expression vector pMG36e-H5LL is successfully constructed.

The basement membrane is a specialized extracellular matrix between the epithelium and the mesenchyme. In stratified epithelium, only the basal cells in contact with the basement membrane exhibit the apical-basal polarity, whereas the epithelial cells do being not in contact with the basement membrane do not exhibit the apical-basal polarity. The basement membrane plays an important role in epithelial cell polarization. It is an important extracellular matrix (ECM) structure in the multicellular organisms, is situated between the epithelium and the mesenchyme, and is produced jointly by the epithelial cells and mesenchymal cells. Its components mainly include Laminin, type Ⅳ collagen (Col-Ⅳ), nidogen(NDG), and heparan sulfate proteoglycans (HSPG), and each component plays the different role in influencing the epithelial cell polarity. The network scaffold formed by Col-Ⅳ and Laminin is the main structure of the basement membrane, and the integrity of the structure affects the epithelial cell polarization. This review summarizes the composition and structure of the basement membrane, focuses on its role in epithelial cell polarization and its mechanism, and compiles the current status of biomimetic basement membrane materials that promotes the epithelial cell polarization, and provides the theoretical foundation for further exploration of the establishment and maintenance of epithelial cell polarity.

Objective To discuss the protective effect of velvet antler peptide （VAP） in the osteoporosis （OP） model rats，and to clarify the possible mechanism. Methods Sixty 12-week-old SD rats were randomly divided into control group， model group， positive drug group （treated with 1 mg·kg^－1·d^－1 of alendronate sodium by gavage）， low dose of VAP group （treated with 100 mg·kg^－1·d^－1 VAP), medium dose of VAP group （treated with 200 mg·kg^－1·d^－1 VAP), and high dose of VAP group （treated with 300 mg·kg^－1·d^－1 VAP), and there were ten rats in each group. Except for control group，the rats in the other groups were injected with dexamethasone （2 mg·kg^－1) to replicate the OP rat model， while the rats in control group were injected with the equivalent volume of saline twice a week for 11 consecutive weeks. Dual-energy X-ray absorptiometry was used to detect the bone mineral density （BMD） of femur tissue of the rats in various groups；enzyme-linked immunosorbent assay （ELISA） method was used to detect the levels of serum calcium （Ca²⁺）， phosphate （P）， osteoprotegerin （OPG）， alkaline phosphatase （ALP）， and osteocalcin （OCN）in serum of the rats in various groups； biochemical method was used to detect the malondialdehyde （MDA） level and superoxide dismutase （SOD） activity in serum of the rats in various groups； HE staining was used to observe the pathomorphology of bone tissue of the rats in various groups； Western blotting method was used to detect the expression levels of silent information regulator 1 （SIRT1）， catalase （CAT）， Runt-related transcription factor 2 （RUNX2）， and forkhead box protein O1 （FOXO1） proteins in bone tissue of the rats in various groups. Results Compared with control group， the BMD of femoral tissue of the rats in model group was decreased （P<0.05）； compared with model group， the BMD of femur tissue of the rats in positive drug group， medium dose of VAP group， and high dose of VAP group were increased （P<0.05 or P<0.01）. Compared with control group， the levels of Ca²⁺， P， OPG， and SOD activities in serum of the rats in model group were decreased （P<0.05）， and the levels of ALP， OCN， and MDA were increased （P<0.05）； compared with model group， the level of OPG in serum of the rats in low dose of VAP group was significantly increased（P<0.05），the levels of Ca²⁺， P， OPG， and activities of SOD in serum of the rats in positive drug group， medium dose of VAP group， and high dose of VAP group were significantly increased （P<0.05 or P<0.01）， and the levels of ALP， OCN， and MDA in serum of the rats in positive drug group and different doses of VAP groups were decreased （P<0.05 or P<0.01）. The HE staining results showed that compared with control group， the rats in model group had fewer bone cells and disordered arrangements in the bone tissue， thinner bone trabeculae with large fractures， and an expanded marrow cavity； compared with model group， the rats in positive drug group， medium dose of VAP group， and high dose of VAP group had thicker bone trabeculae arranged more tightly. The Western blotting results showed that compared with control group， the expression levels of SIRT1， CAT， RUNX2， and FOXO1 proteins in bone tissue of the rats in model group were decreased （P<0.05）； compared with model group， the expression levels of SIRT1， CAT， RUNX2， and FOXO1 proteins in bone tissue of the rats in positive drug group， medium dose of VAP group， and high dose of VAP group were significantly increased （P<0.05 or P<0.01）. Conclusion VAP has the protective effect against OP in the rats， and its mechanism may be related to mediating the antioxidant stress action through the SIRT1/FOXO1 signaling pathway.

Objective To analyze the clinical presentations and diagnostic and treatment process of one patient with autoimmune encephalitis（AE） with double positive anti-N-methyl-D-aspartate receptor （NMDAR） and anti-γ-aminobutyric acid B receptor （GABA_BR） secondary to herpes simplex virus encephalitis（HSVE），and to improve the clinicians’ awareness of this disease. Methods The clinical data of one AE patient with double positive anti-NMDAR and anti-GABA_BR secondary to HSVE were collected， the diagnostic and therapeutic processes were summarized， and the relevant literatures were reviewed. Results The patient， a 36-year-old male， developed a headache followed by limb convulsions， and progressed to disturbed consciousness. After admission， the routine biochemistry of the cerebrospinal fluid （CSF） was abnormal， and the herpes simplex virus-1 （HSV-1） IgG antibody showed positive in the CSF； both CSF and serum tests for NMDAR antibodies were positive； the head magnetic resonance imaging （MRI） results showed abnormal signals in the right occipital white matter， leading to the diagnosis of HSVE secondary to anti-NMDAR encephalitis. Several months later， the patient experienced psychiatric behavior abnormalities， cognitive dysfunction， and sleep disorders， and both the serum NMDAR and GABA_BR antibodies showed positive results， prompting the diagnosis of HSVE secondary anti-NMDAR encephalitis and anti-GABA_BR encephalitis. After treatment with steroid pulse therapy and intravenous immunoglobulin （IVIG）， the patient’s condition was improved and the patient was discharged. At one-year follow-up， the patient’s psychiatric symptoms had completely resolved， leaving mild cognitive impairment. Conclusion If the clinical symptoms of the patients recovering from antiviral treatment for HSVE is worsened， secondary AE should be highly suspected；it is important to complete autoimmunity antibody testing as soon as possible for the early diagnosis and treatment to improve the prognosis of the patient.

Objective To discuss the protective effect of gingerone on the hippocampal neuron HT22 cells after oxygen-glucose deprivation/reoxygenation （OGD/R），and to clarify the related mechanism. Methods The HT22 cells were cultured， and the OGD/R cell injury model was established by setting the gradient of OGD/R time. The HT22 cells were divided into control group，OGD/R group， OGD/R+1 μmol·L^－1 gingerone group， OGD/R + 10 μmol·L^－1 gingerone group， OGD/R+100 μmol·L^－1 gingerone group，and OGD/R+0.2% dimethyl sulfoxide（DMSO） group.The viability of the cells in various groups was detected by CCK-8 assay； the survival rates of the cells in various groups were calculated to determine the optimal drug concentration of gingerone. The cells were divided into control， OGD/R group， OGD/R+ gingerone， and OGD/R+gingerone+nuclear factor erythroid-2-related factor 2（Nrf2） inhibitor（ML385） groups.The cells in OGD/R + gingerone group were treated with gingerone for 4 h before OGD treatment for 8 h followed by reoxygenation for 8 h， and the cells in OGD/R+gingerone+ML385 group were treated with 10 μmol·L^－1 ML385 for 6 h before gingerone treatment. The viability of the cells in various groups was detected by CCK-8 assay；the expression levels of Nrf2， heme oxygenase-1 （HO-1）， B-cell lymphoma-2 （Bcl-2）， and Bcl-2-associated X protein （Bax） proteins in the cells in various groups were detected by Western blotting method；the activity of superoxide dismutase （SOD） and the level of malondialdehyde （MDA） in the cell culture supernatant in various groups were detected by enzyme-linked immunosorbent assay （ELISA） method. Results Compared with control group， the survival rate of the HT22 cells was below 50% after treated with OGD for 8 h and reoxygenation for 8 h， so the HT22 cell OGD/R model was established by treated with OGD for 8 h and reoxygenation for 8 h. Compared with OGD/R group，the survival rates of the cells in OGD/R+different doses of gingerone groups were increased to various extents， and the survival rate of the cells in OGD/R+ 100 μmol·L^－1 gingerone group was significantly increased （P<0.01）； so 100 μmol·L^－1 gingerone was used for the subsequent experiment. Compared with control group， the viability of the cells in OGD/R group was significantly decreased （P<0.01）， and the expression levels of Nrf2， HO-1， and Bax proteins in the cells were significantly increased （P<0.01）， while the expression level of Bcl-2 protein in the cells was significantly decreased （P<0.05）， and the SOD activity in the cell culture supernatant was significantly decreased （P<0.01）， and the level of MDA was significantly increased （P<0.01）； compared with OGD/R group， the viability of the cells in OGD/R + gingerone group was significantly increased （P<0.01）， and the expression levels of Nrf2， HO-1， and Bcl-2 proteins in the cells were significantly increased （P<0.05 or P<0.01）， while the expression level of Bax protein in the cells was decreased（P<0.05）， the SOD activity in the cell culture supernatant was significantly increased （P<0.01）， and the level of MDA was significantly decreased （P<0.01）； compared with OGD/R + gingerone group， the viability of the cells in OGD/R + gingerone + ML385 group was significantly decreased （P<0.01）， and the expression levels of Nrf2， HO-1， and Bcl-2 proteins were significantly decreased （P<0.01）， while the expression level of Bax protein in the cells was significantly increased （P<0.01），the SOD activity in the cell culture supernatant was significantly decreased （P<0.01）， and the level of MDA was significantly increased （P<0.05）. Conclusion Gingerone alleviates the oxidative stress damage，and thereby plays an inhibiory effect on the apoptosis of the HT22 neurons by activating the Nrf2/HO-1 signaling pathway after OGD/R.

Objective To discuss the effect of downregulating of high mobility group box protein 2 （HMGB2） expression on the biological behavior of the liver cancer cells and the epithelial-mesenchymal transition （EMT） process， and to clarify its mechanism. Methods The human liver cancer LM3 cells at logarithmic growth phase were divided into negative control group and HMGB2 RNA interference group （HMGB2 siRNA group）；the cells in two groups were transfected with RNA oligonucleotides（RNA oligos） with irrelevant sequences and RNA oligos designed to knock down HMGB2， and the Lipofectamine 2000 was regarded as the vector.The expression levels of HMGB2 mRNA and protein in the cells in two groups were detected by real-time fluorescence quantitative PCR （RT-qPCR） and Western blotting methods； cell scratch assay and Transwell chamber assay were used to detect the migration and invasion abilities of the cells in two groups； the expression levels of E-cadherin， N-cadherin， and Vimentin proteins and protein kinase B （AKT）/mammalian target of rapamycin （mTOR） pathway related proteins in the cells in two groups were detected by Western blotting method. Results Compared with negative control group， the expression levels of HMGB2 mRNA and protein in the cells in HMGB2 siRNA group were significantly decreased （P<0.05），the cell scratch healing rate was significantly decreased （P<0.01），the number of invasion cells was significantly decreased （P<0.01），and the expression level of E-cadherin protein in the cells was significantly increased （P<0.01）， while the expression levels of N-cadherin， Vimentin， mTOR， AKT， and phosphorylated AKT （p-AKT） proteins in the cells were significantly decreased （P<0.05 or P<0.01）. Conclusion Downregulating the expression of HMGB2 can reduce the migration and invasion abilities of the liver cancer LM3 cells and inhibit the EMT，and its mechanism may be related to regulating the expression of the AKT/mTOR pathway related proteins.

Objective To analyze the imaging performance and clinical diagnosis and treatment process of one patient with peri-gallbladder fluid caused by the posterior duodenal perforation primarily diagnosed by ultrasound， and to provide the clinical diagnostic evidence for this disease. Methods The clinical data， laboratory examination， gastroscope performance， and imaging performance of one patient with peri-gallbladder fluid caused by the posterior duodenal perforation was collected. The process of diagnosis and treatment was recorded and followed up. The related literatures were reviewed to analyze the clinical characteristics and imaging performances of the posterior duodenal perforation. Results The patient was a 50-year-old male who had constant dull pain in the upper right abdomen for over 20 d， without obvious cause and intensified after meals， accompanied by radiculalgia in the right waist and back.The ultrasonic examination at a local hospital showed an occupying lesion in the gallbladder， and the patient came to our hospital for the further diagnosis and treatment. On the day of admission， the ultrasound examination showed poor fasting gallbladder filling， continuous and uniformly thickened gallbladder wall.The multiple strong echoes were observed in the gallbladder cavity， and the chaotic distributed hypoechoic and anechoic areas could be seen around the gallbladder， extending to the back of the duodenal bulb， and connected with the bulb by a narrow strip of gaseous shadow. The ultrasound results showed that there was perforation of the posterior wall of the duodenal bulb， leading to pericholecystic fluid accumulation， with poor gallbladder filling due to compression， and chronic inflammation of the gallbladder wall associated with gallstones. The enhanced computed tomography （CT） results clearly showed the perforation site in the posterior wall of the duodenal bulb， and the CT conclusion was consistent with the ultrasound conclusion. The gastroscope results showed a large deep ulcer on the posterior wall of the duodenal bulb， and it confirmed to be diagnosed as the peri-gallbladder fluid caused by ulcer perforation. A gastric tube and jejunal nutrition tube were inserted， and the symptomatic treatments such as anti-inflammation， analgesia， and acid suppression were carried out. The ultrasound reexamination within 3 months of treatment showed the peri-gallbladder fluid was gradually decreased and the gallbladder cavity was gradually filled.Since the patient’s gallbladder inflammation symptoms persisted，the cholecystectomy was performed， and it was pathologically diagnosed as chronic cholecystitis complicated with gallstones. Conclusion The posterior duodenal perforation is often misdiagnosed or missed due to atypical clinical manifestations.The imaging examination can assist in early clinical diagnosis. When the presence of abdominal gas does not interfere with observation， the ultrasound can serve as the first choice and effective method for the diagnosis of gastrointestinal perforation.

Objective To prepare the polypropylene carbonate（PPC）/polybutylene succinate（PBS） biofilm that mimics the structure of human bone periosteum，and to evaluate its physicochemical properties and biological characteristics. Methods The PPC/PBS biofilm was prepared by salting-out method. ¹H nuclear magnetic resonance spectra（¹HNMR） was used to observe the spectral absorption peak of PPC， PBS， and PPC/PBS，and the chemical structure changes of PPC， PBS， and PPC/PBS were analyzed； the ultramorphology of the PPC/PBS biofilm was observed by scanning electron microscope （SEM）； and the physicochemical properties， including porosity， Young’s modulus， fracture strength， fracture elongation， and static contact angle of the PPC/PBS biofilm were measured by GPC membrane permeation chromatography. The osteoblasts of the primary SD rat were isolated， cultured， and purified， and divided into control group （without biofilm）， BME-10X collagen membrane group， and PPC/PBS biofilm group. SEM was used to observe the adhesion and growth of the osteoblasts in various groups； cell counting was performed to detect the number of osteoblasts；alkaline phosphatase （ALP） assay kit was used to the differentiation of the osteoblasts； rabbit muscle degradation experiment was used to evaluate the degradation performance of the PPC/PBS biofilm in vivo. Results The PPC/PBS biofilm had a dual-layer structure consisting of a smooth surface layer and a rough surface layer， with a total thickness of approximately 0.5 mm and an average pore size of about 120 μm；the porosity was approximately 77.4%， the Young’s modulus was approximately 38.1 MPa， the fracture strength was approximately 1.22 MPa， the fracture elongation was approximately 7.4%， and the contact angle on the rough surface was 85°， while the contact angle on the smooth surface was 57°. The SEM observation results showed that fewer cells adhered to the surface of the PPC/PBS biofilm 1 d after culture； 3， 7， and 14 d after culture， a large number of osteoblasts were observed to adhere and grow on the surface of the biofilm， with cell protrusions attached to the film and cell bodies distributed in the pores. Compared with control group， the numbers of osteoblasts attached to the surface of the materials in BME-10X collagen membrane group and PPC/PBS biofim group were decreased after cultured for 1， 3， 7， and 14 d （P<0.05）. Compared with BME-10X collagen membrane group， the number of the osteoblasts attached to the surface of the materials in PPC/PBS biofilm group was increased after cultured for 1， 3， 7， and 14 d （P<0.05）. Compared with control group， the ALP levels in the osteoblasts attached to the surface of the materials in BME-10X collagen membrane group and PPC/PBS biofilm group were significantly decreased after cultured for 1， 3， 7， and 14 d （P<0.01）. Compared with BME-10X collagen membrane group， the ALP level in the osteoblasts attached to the surface of the materials in PPC/PBS biofilm group was significantly increased after cultured for 1 and 3 d （P<0.01）. The degradation experiment results showed that compared with 0 week after degradation， the weight loss rate and weight loss rate of number-avarage molecular of the PPC/PBS biofilm were increased 4 weeks after degradation （P<0.05）， while the fracture strength and fracture elongation were significantly decreased（P<0.05 or P<0.01）. From the 2nd week after degradation，the number of microstructures on rough surface of the PPC/PBS biofilm was gradually increased， and the pore sizes was ranged from 0 to 10 μm； by 26 weeks after degradation，the number of micro-porous structures was evenly distributed on rough surface of the PPC/PBS biofilm； by 4 weeks after degradation， the smooth surface of the biofilm showed exfoliation changes， but no micro-porous structures were observed； by 12 weeks after degradation， a small number of micro-porous changes were observed on the smooth surface， and the pore diameters were less than 5 μm； by 26 weeks after degradation， the number of micro-porous structures on the smooth surface was increased，and the pore diameters were less than 10 μm. Conclusion The structure of PPC/PBS biofilm is similar to that of human bone outer membrane，with a double-layer structure，good hydrophilicity，dense smooth surface，high porosity of rough surface，good biocompatibility，and slow degradation；it is an ideal guided regeneration biofilm.

Objective To discuss the potential mechanism of Panax quinquefolium triolsaponins （PQTS） in the occurrence and development of ischemic stroke by using network pharmacology and molecular docking technique. Methods The potential targets of PQTS acing on IS were obtained through Swiss Target Prediction Database， Encyclopedia of Traditional Chinese Medicine（ETCM） Database， SEA Search Server Database， DisGeNET Database， and so on； the protein-protein interaction （PPI） network diagram of the key potential targets was established by STRING Database and Cytoscape 3.9.1 software；the core tagets of PQTS acting on IS were got by topology network analysis； Gene Ontology （GO） function and Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathway enrichment analysis were used to analyze the potential targets through DAVID online analysis website； the PQTS-target-signaling pathway network was constructed by Cytoscape 3.9.1 software and the topology network analysis was used to obtain the potential main active compositions； AutoDock Vina software was used to verify the molecular docking between the active ingredients and core targets. Results There were 122 potential targets of PQTS acting on IS； the GO function enrichment analysis was mainly included the regulation of apoptosis， intracellular signal transduction process， and regulations of extracellular substances by cells； the KEGG function analysis included the interleukins signaling pathways， phosphatidylinositol 3 kinase/protein kinase B （PI3K/Akt） signaling pathway and phosphatidylinositol 3 kinase-protein kinase B-mammalian target of rapamycin （PI3K-Akt-mTOR） signaling pathway. The molecular docking analysis results showed that pseudo-ginsenoside F11， 20（S）-protopanaxatriol， ginsenoside Rg1， ginsenoside Rh1， pseudo-ginsenoside RT5， and ginsenoside Re could form the stable conformations with signal transducer and activator of transcription 3（STAT3），phosphatidylinositol 3-kinases catalytic suburit α（PIK3CA），epidermal growth factor receptor （EGFR）， and mitogen-activated protein kinase 14（MAPK14） with lower binding energy. Conclusion The protective effect of PQTS on IS may be related to the STAT3， PIK3CA， EGFR， MAPK14， and PI3K/Akt signaling pathways.

Objective： To analyze the clinical performance and diagnosis process of the autoimmune encephalitis （AE） patients with positive double antibodies in the cerebrospinal fluid（CSF）， and to provide the references for the diagnosis and treatment of such patients. Methods The clinical manifestations， magnetic resonance imaging （MRI） of the head， electroencephalogram （EEG）， CSF characteristics， and prognosis of one patient with AE positive for anti-glutamic acid decarboxylase （GAD） 65 and anti-γ-aminobutyric acid B receptor （GABA_BR） double antibodies in the CSF were retrospectively analyzed，and the literatures were reviewed. Results The patient was a 47-year-old male with subacute onset and progressively aggravated symptoms， mainly presenting with headaches and episodic convulsions， and blurred consciousness.The MRI results of the head suggested that the lesions were located on the right side of the cerebral falx in the frontal and parieto-occipital lobes；the CSF was positive for the anti-GAD65 and anti-GABA_BR double antibodies， and the EEG results showed the abnormal spike and slow waves， so the patient was diagnosed as AE. After anti-inflammatory and other symptomatic treatments， the patient was gradually improved and was discharged；the patient was given continuous oral corticosteroid treatment， but after 5 months， the patient was relapsed with acute onset， presenting with convulsion accompanied by drooling from the corner of the mouth. The head MRI results showed there was an abnormal high signal in the right temporal lobe. After corticosteroid treatment， the patient was improved. Conclusion The AE patients with positive double antibodies in CSF are more likely to relapse. Steroid anti-inflammatory treatment is effective. When intracranial lesions are located in the frontal and parieto-occipital lobes， it should be considered the possibility of symptoms such as convulsions. It is necessary to complete the EEG and the other inspections as soon as possible.

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 discuss the potential categories of adolescents’ negative physical intentions， and to further analyze the relationships between different potential categories and suicidal ideation and non-suicidal self-injury of adolescents’ negative physical intentions. Methods In Changchun City of Jilin Province，4 middle schools were randomly selected， and 2 or 3 teaching classes were selected from each grade by using cluster randomly sampling method，and 1 459 students were regarded as the survey subjects， Beck Scale for Suicide Ideation Chinese Version（BSI-CV）， Adolescent Self Harm Scale （ASHS）， and Negative Physical Self Scale （NPSS） were used for the on-the-spot questionnaire survey， and the results were analyzed by latent profile analysis，χ² test，and Logistic regression analysis. Results The adolescents’ negative physical intentions were divided into body satisfaction（65.2%，951 persons），emaciated body dissatisfaction（13.4%，195 persons） and obese appearance dissatisfaction（21.4%，313 persons）.The Logistic regression analysis results showed that only-child（OR=2.43，95%CI：1.21－4.89，P<0.05）， bullying experience（OR=2.43， 95%CI：2.19－5.72，P<0.05），emaciated body dissatisfaction（OR =5.42， 95%CI：2.66－11.05，P<0.01），and obese appearance dissatisfaction（OR=9.34， 95%CI：5.18－16.83，P<0.01） were the risk factors for the adolescents’ suicidal ideation； girls（OR=2.35，95%CI：1.49－3.71，P<0.01），single-parent family（OR=1.99，95%CI：1.11－3.59，P<0.05），bullying experience（OR=5.26， 95%CI：3.42－8.08，P<0.01），emaciated body dissatisfaction（OR=2.34， 95%CI：1.21－4.53，P<0.05），and obese appearance dissatisfaction（OR=5.24， 95%CI：3.31－8.28，P<0.01）were the risk factors for the non-suicidal self-injury of the adolescents. Conclusion The negative physical intentions of the adolescents have heterogeneous；emaciated body dissatisfaction and obese appearance dissatisfaction are the risk factors for the suicidal ideation and non-suicidal self-injury.

Objective： To discuss the effect of human ceramide 1-phosphate transfer protein （CPTP） on the biological behavior of oral squamous cell carcinoma HSC-3 cells，and to clarify its related mechanism. Methods The HSC-3 cells were cultured in vitro and divided into control group and experiment group，and the cells in two groups were transfected with pFlag-CMV4 and pFlag-CPTP plasmids，respectively；the cells stably transfected with CPTP were constructed by using resistance screening method. Western blotting and immunofluorescence methods were used to detect the expression levels of CPTP protein in cells in two groups； clone formation experiment and CCK-8 assay were used to detect the numbers of clone formation and proliferation activities of cells in two groups； cell scratch experiment was used to detect the scratch healing rates of cells in two groups； Transwell chamber experiment was used to detect the numbers of invasion cells in two groups. The mice were randomly divided into control group and experiment group， and the mice were injected with the HSC-3 cells stably transfected with pFlag-CMV4 and the HSC-3 cells stably transfected with pFlag-CPTP respectively to construct the subcutaneous transplanted tumor models in the mice.The volumes and weights of transplant tumors of the mice in two groups were detected. The enrichment analysis on CPTP differentially expressed genes in head and neck squamous cell carcinoma （HNSCC） cells was analyzed by using bioinformatics method；real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of p53， thrombospondin-1 （THBS1）， and cyclin G2 （CCNG2） mRNA in cells in two groups. Results Compared with control group， the expression amount of CPTP protein in the cells in experiment group was increased.Compared with control group， the number of clone formation in the cells in experiment group was significantly increased （P<0.01）， the proliferation activity and scratch healing rate of the cells were significantly increased （P<0.05 or P<0.01）， and the number of invasion cells was increased （P<0.01）. After injecting tumor cells for 2，3，and 4 weeks， compared with control group， the volumes of the transplanted tumor of the mice in experiment group were increased （P<0.05 or P<0.01）；after injecting tumor cells for 4 weeks，compared with control group，the weight of transplanted tumor of the mice in experiment group was increased（P<0.05）. The bioinformatics analysis results showed that the role of CPTP in the HNSCC tissue may be mediated through the signaling pathways such as p53. Compared with control group， the expression levels of p53， THBS1， and CCNG2 mRNA in the cells in experiment group were significantly decreased （P<0.01）. Conclusion Over-expression of CPTP can promote the proliferation， migration， invasion， and tumorigenesis of the HSC-3 cells. CPTP promotes the growth of the oral squamous cell carcinoma HSC-3 cells by inhibiting the p53 signaling pathway.

Objective To discuss the resistance and regeneration effects of long non-coding RNA （lncRNA） GPRC5D-AS1 on the muscle atrophy of myocytes in the mice induced by dexamethasone， and to clarify its mechanism. Methods The C2C12 cells at logarithmic growth phase were selected. The survival rates of the cells after treated with 0， 25， 50， 75， 100， 200 and 400 mg·L^－1 dexamethasone for 24， 48 and 72 h were detected， which was used to select the optimal dose and time to induce the muscle atrophy model.The expression level of lncRNA GPRC5D-AS1 in the C2C12 cells was detected by real-time fluroscence quantitative PCR（RT-qPCR） method to validate the cell model. The C2C12 cells were divided into normal group， control group，lncRNA GPRC5D-AS1-NC group，and lncRNA GPRC5D-AS1-OE group. The expression levels of lncRNA GPRC5D-AS1 in the cells in various groups were detected by RT-qPCR method. The levels of reactive oxygen species （ROS）， glutathione peroxidase 4 （GPX4）， ferrous ion （Fe²⁺） and malondialdehyde （MDA） in the cells in various groups were detected by flow cytometry and enzyme-linked immunosorbent assay （ELISA） method； the ultrastructural changes of mitochondria in the cells in various groups were observed under transmission electron microscope； the expression of myogenic differentiation protein （MyoD） in the cells in various groups was detected by immunofluorescence； the expression levels of ferroptosis-related proteins in the cells in various groups were detected by Western blotting method. Results The best modeling effect was achieved with 100 mg·L^－1 dexamethasone at 48 h. Compared with normal C2C12 cells， the expression level of lncRNA GPRC5D-AS1 in the C2C12 cells after treated with 100 mg·L^－1 dexamethasone for 48 h was decreased （P<0.05）， confirming that lncRNA GPRC5D-AS1 had a regulatory role in the muscle atrophy model. The RT-qPCR results showed that compared with normal group， the expression level of lncRNA GRPC5D-AS1 in the cells in model group was decreased （P<0.05）； compared with model group and lncRNA GRPC5D-AS1-NC group， the expression level of lncRNA GRPC5D-AS1 in the cells in lncRNA GRPC5D-AS1-OE group was increased （P<0.05）. The flow cytometry results showd that compared with normal group， the ROS level in the cells in model group was increased （P<0.05）； compared with model group and lncRNA GRPC5D-AS1-NC group， the ROS level in the cells in lncRNA GRPC5D-AS1-OE group was decreased （P<0.05）. The ELISA results showed that compared with normal group， the levels of Fe²⁺ and MDA in the cells in model group were increased （P<0.05）， and the level of GPX4 was decreased （P<0.05）； compared with model group and lncRNA GRPC5D-AS1-NC group， the levels of Fe²⁺ and MDA in the cells in lncRNA GRPC5D-AS1-OE group were decreased （P<0.05）， and the level of GPX4 was increased （P<0.05）. The transmission electron microscope results showed that the cells in normal group were round and normal in size with rich cilia on the membrane， and the mitochondria had a normal morphology； the cells in model group had less mitochondria and granulation in the cytoplasm， with blurred and swollen mitochondrial structures and typical ferroptosis-related manifestations； the number of mitochondria in the cells in lncRNA GRPC5D-AS1-OE group was increased，and the granulation in cytoplasm of the cells was decreased， and showed clearer mitochondrial structures. The immunofluorescence detection results showed that compared with normal group，the expression of MyoD in the cells in model group was decreased and the cells exhibited muscle atrophy； compared with model group and lncRNA GRPC5D-AS1-NC group， the expression of MyoD in the cells in lncRNA GRPC5D-AS1-OE group was increased， and the cells exhibited less muscle atrophy and more regeneration of myocytes. The Western blotting results revealed that compared with the normal group， the expression level of long-chain acyl-coenzyme A synthase （ACSL）4 protein in the cell in model group was increased （P<0.05）， and the expression levels of solute carrier family 7 member 11 （SLC7A11） and GPX4 proteins were decreased （P<0.05）； compared with model group and lncRNA GRPC5D-AS1-NC group， the expression level of ACSL4 protein in the cells in lncRNA GRPC5D-AS1-OE group was decreased （P<0.05）， and the expression levels of SLC7A11 and GPX4 proteins were increased （P<0.05）. Conclusion LncRNA GPRC5D-AS1 can protect the ferroptosis in the myocytes induced by dexamethasone and promote the repairment and regeneration of the cells，and its mechanism is related to regulating the SLC7A11/GPX4/ACSL4 signaling pathway.

Objective To discuss the effect of Cadherin-17 on the proliferation and apoptosis of the colorectal cancer （CRC） cells，and to clarify its possible mechanism. Methods The Cadherin-17 gene over-expression and small interference plasmids were constructed and packaged as the lentivirus and transfected into the SW480 cells to construct the stable transfection strain of over-expression and interference virus. The expression levels of Cadherin-17 mRNA and protein in the cells were detected by real-time fluorescence quantitative PCR （RT-qPCR） and Western blotting methods， and the transfection efficiency was verified and the stable transfection strain was identified. The SW480 cells were divided into control group， empty vector group， Cadherin-17 over-expression plasmid （OV-Cadherin-17） group and Cadherin-17 small interference plasmid （si-Cadherin-17） group. The activities of cells in various groups were detected by CCK-8 assay；the apoptotic rates of cells in various groups were detected by flow cytometry； the expression levels of Cadherin-17，B-cell lymphoma-2 （Bcl-2）， Bcl2-associated X （Bax）， cytochrome c （Cyt-c） and cysteinyl aspartate specific proteinase-3 （Caspase-3），and the phosphatidylinosital-3-kinase/protein kinase B/mamalian target of repamycin （PI3K/AKT/mTOR） signaling pathway-related proteins in the cells in various groups were detected by Western blotting methods. The cells were treated with PI3K inhibitor LY294002 and divided into control group， LY294002 group， OV-Cadherin-17+LY294002 group，and si-Cadherin-17+LY294002 group； the proliferation activities and apoptotic rates of cells in various groups and the expression levels of Bcl-2，Bax，Cyt-c，Caspase-3 and the expression levels of PI3K/AKT/mTOR signaling pathway-related proteins in the cells in various groups were detected. Results The RT-qPCR and Western blotting results showed that the OV-Cadherin-17 and si-Cadherin-17 transfection and stable transfection stain were successfully constructed.Compared with control group， the proliferation ability of the cells in OV-Cadherin-17 group was increased （P<0.01）， the apoptotic rate was decreased （P<0.01）， the expression levels of Bax and Caspase-3 proteins in the cells were decreased （P<0.01）， the expression levels of Bcl-2 and Cyt-c proteins in the cells were increased （P<0.01）， and the expression levels of phosphorylated PI3K（p-PI3K），phosphorylated AKT（p-Akt）， and phosphorylated mTOR（p-mTOR） proteins were increased （P<0.01）； the proliferation abilities of the cells in si-Cadherin-17 and LY294002 groups were decreased （P<0.01）， the apoptotic rates were increased （P<0.01）， the expression levels of Bax and Caspase-3 proteins in the cells were increased （P<0.01）， the expression levels of Bcl-2 and Cyt-c proteins in the cells were decreased （P<0.01），and the expression levels of p-PI3K， p-AKT， and p-mTOR proteins in the cells were decreased （P<0.01）； compared with LY294002 group， the proliferation ability of the cells in OV-Cadherin-17+LY294002 group was increased （P<0.01）， the apoptotic rate was decreased （P<0.01）， the expression levels of Bax and Caspase-3 proteins in the cells were decreased （P<0.01）， the expression levels of Bcl-2 and Cyt-c proteins in the cells were increased （P<0.01）， the expression levels of p-PI3K， p-AKT， and p-mTOR proteins were increased （P<0.01）， the proliferation activity of the cells in si-Cadherin-17+LY294002 group was decreased （P<0.01）， the apoptotic rate was increased （P<0.01）， the expression levels of Bax and Caspase-3 proteins in the cells were increased （P<0.01）， the expression levels of Bcl-2 and Cyt-c proteins in the cells were decreased （P<0.01），and the expression levels of p-PI3K， p-AKT， and p-mTOR proteins were decreased （P<0.01）. Conclusion Cadherin-17 can promote the proliferation and inhibit the apoptosis of the CRC cells， and its mechanism may be related to the activition of PI3K/AKT/mTOR signaling pathway regulated by Cadherin-17.

Objective To analyze the efficacy of anterior cervical Hybrid surgery and posterior cervical expansive open-door laminoplasty （EODL） in the treatment of multilevel cervical spondylotic myelopathy， and to discuss the selection of surgical methods for the patients with multilevel cervical spondylotic myelopathy. Methods The retrospective analysis was conducted of 70 patients with multilevel cervical spondylotic myelopathy who underwent surgery at Affilated Beijing Traditional Chinese Medicine Hospital of Capital Medical University from July 2017 to July 2020. Based on the different surgical methods， the patients were divided into anterior group （n=35 ）and posterior group（n=35）. The patients in anterior group underwent Hybrid surgery ［anterior cervical discectomy and fusion （ACDF） combined with artificial cervical disc replacement （ACDR）］，and the patients in posterior group underwent EODL. The hospitalization time， operation time， intraoperative blood loss， and postoperative drainage volume of the patients in two groups were recorded； the efficacy was evaluated by Japanese orthopaedic association （JOA） score， JOA improvement rate， neck disability index （NDI）， visual analogue scale （VAS） for pain， and postoperative satisfaction score； the complications of the patients in two groups after surgery were recorded. Results Compared with posterior group， the intraoperative blood loss， postoperative drainage volume， hospitalization time， and operation time of the patients in anterior group were significantly decreased （P<0.01）， and the preoperative score had no significant difference （P>0.05）. At the final follow-up after surgery， compared with posterior group， the JOA score and JOA improvement rate of the patients in anterior group were significantly increased （P<0.01）， and the NDI score and VAS score were significantly decreased （P<0.01）.Compared with before surgery， the JOA scores of the patients in two groups at the final follow-up after surgery were increased （P<0.01）， and the NDI and VAS scores were significant decreased （P<0.01）. The postoperative satisfaction of the patients in two groups was high based on the postoperative satisfaction score.There was no significant difference in the incidence of postoperative complication of the patients between two groups （P>0.05）. Conclusion Both the anterior cervical Hybrid surgery and EODL achieve the satisfactory results in the treatment of multilevel cervical spondylotic myelopathy. Hybrid surgery has the advantages of less bleeding and shorter surgery time， and the most suitable surgical method should be chosen clinically based on the actual situation of the patients.

Objective To discuss the effect of honeysuckle extract on the proliferation and apoptosis of airway smooth muscle cells （ASMCs） in the asthmatic mice，and to clarify the related mechanism. Methods The asthma models of mice were constructed， and the primary ASMCs were isolated and identified. The M2 polarization of macrophages RAW264.7 was induced and identified. The RAW264.7 cells were divided into control group， low， medium and high doses of honeysuckle extract groups. The RAW264.7 cells in control group were induced M2 polarization and co-cultured with ASMCs. The RAW264.7 cells in low， medium， and high doses of honeysuckle extract groups were treated with different concentrations （50， 100， and 200 mg·L^－1） of honeysuckle extract for 1 h and then treated with interleukin-4 （IL-4） （60 μg·L^－1） for 6 h. Then the RAW264.7 cells and ASMCs were co-cultured for 24 h. The percentages of CD86 and CD206 positive cells in various groups were detected by flow cytometry； the levels of interleukin-10（IL-10） in culture supernatant of the ASMCs in various groups were detected by enzyme-linked immunosorbent assay （ELISA） method； the proliferation activities of cells in various groups were detected by MTT assay； the apoptotic rates of ASMCs in various groups were detected by flow cytometry. Results The morphology of the cells and immunofluorescence staining results proved that the extracted cells were the ASMCs.The M2 polarization identification results showed that the RAW264.7 cells were induced into the M2 macrophages.Compared with control group， the percentages of CD86 positive cells in the ASMCs in low， medium and high doses of honeysuckle extract groups were significantly increased （P<0.05），while the percentages of CD206 positive cells were decreased（P<0.05）；the IL-10 levels in culture supernatant of the ASMCs and the proliferation activities of ASMCs were significantly decreased （P<0.05），and the apoptotic rates of the ASMCs were increased（P<0.05）. Compared with low dose of honeysuckle extract group， the percentages of CD86 positive cells in medium and high doses of honeysuckle extract groups were significantly increased（P<0.05）， while the percentages of CD206 positive cells were decreased（P<0.05）；the IL-10 levels in the culture supernatant and proliferation activities of the cells were significantly decreased （P<0.05），and the apoptotic rates of the ASMCs were increased（P<0.05）.Compared with medium dose of honeysuckle extract group， the percentages of CD86 positive cells in high dose of honeysuckle extract group was significantly increased（P<0.05）， while the percentage of CD206 positive cells was decreased（P<0.05）； the IL-10 levels in the culture supernatant and proliferation activities of the cells were significantly decreased （P<0.05），and the apoptotic rates of the ASMCs were increased（P<0.05）. Conlusion： Honeysuckle extract can inhibit the proliferation and promote the apoptosis of the ASMCs in the asthmatic mice by inhibiting the M2 polarization of the macrophages.

Objective To discuss the protective effect of fructus mume total flavonoids （FMF） on the injury of the SH-SY5Y cells induced by 1-methyl-4-phenylpyridinium（MPP⁺） through regulating microRNA （miR）-145-3p expression，and to clarify the mechanism. Methods The SH-SY5Y cell model of Parkinson’s disease （PD） was established by MPP⁺ induction. The optimal intervention concentration and action time of MPP⁺ and FMF were screened out by CCK-8 assay. The cells were divided into control group （untreated）， model group （treated with 500 μmol·L^－1 MPP⁺ for 24 h）， MPP⁺+mimics group （transfected with miR-145-3p mimics）， MPP⁺+NC group （transfected with miR-145-3p NC）， MPP⁺+FMF group treated with 0.25 g·L^－1 FMF for 24 h）， MPP⁺+mimics+FMF group （transfected with miR-145-3p mimics and treated with 0.25 g·L^－1 FMF for 24 h），and MPP⁺+NC+FMF group （transfected with miR-145-3p NC，and （treated with 0.25 g·L^－1 FMF for 24 h）. CCK-8 assay was used to detect the proliferation abilities of the cells in various groups； Annexin Ⅴ-FITC/PI staining was used to the detect the apoptotic rates of the cells in various groups； transmission electron microscope was used to observe the ultrastructure morphology of mitochondrial autophagy in the cells in various groups； real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of miR-145-3p in the cells in various groups； Western blotting method was used to detect the expression level of Beclin-1 protein and the ratio of LC3-Ⅱ/ LC3-Ⅰ in the cells in various groups. Results After treated with different concentrations of MPP⁺， the proliferation ability of the cells was significantly decreased（P<0.01）， and the PD cell model was successfully constructed. The optimal concentrations and action time of MPP⁺ and FMF intervention were 500 μmol·L^－1， 24 h， and 0.25 g·L^－1，24 h， respectively.The CCK-8 assay results showed that the proliferation ability of the cells in model group was significantly lower than that in control group （P<0.01）； the proliferation abilities of the cells in MPP⁺+mimics group and MPP⁺+FMF group were significantly higher than that in model group（P<0.01）； the proliferation ability of the cells in MPP⁺+mimics+FMF group was significantly higher than that in MPP⁺+FMF group （P<0.01）. The Annexin V-FITC/PI staining results showed that the apoptotic rate of the cells in model group was significantly higher than that in control group （P<0.01）； the apoptotic rates of the cells in MPP⁺+mimics group and MPP⁺+FMF group were significantly lower than that in model group （P<0.01）； the apoptotic rate of the cells in MPP⁺+mimics+FMF group was significantly lower than that in MPP⁺+FMF group （P<0.01）. The transmission electron microscope results showed that the cells in control group showed the uniform and intact mitochondrial morphology， the cells in model group showed the enlarged and irregular mitochondria with vacuolar changes，and the cells in MPP⁺+mimics group and MPP⁺+FMF group showed the improved mitochondrial structure， compared with model group， the autophagosome numbers of the cells in MPP⁺+mimics group and MPP⁺+FMF group were increased； the cells in MPP⁺+mimics+FMF group showed more intact mitochondrial structure， compared with MPP⁺+FMF group， the autophagosome numbers in the cells was increased. The RT-qPCR results showed that the expression level of miR-145-3p in the cells in model group was significantly lower than that in control group （P<0.01）； the expression levels of miR-145-3p in the cells in MPP⁺+mimics group and MPP⁺+FMF group were significantly higher than that in model group （P<0.01）； the expression level of miR-145-3p in the cells in MPP⁺+mimics+FMF group was significantly higher than that in MPP⁺+FMF group （P<0.01）.The Western blotting results showed that the expression level of Beclin-1 protein and the ratio of LC3-Ⅱ/LC3-Ⅰ in the cells in model group were lower than those in control group （P<0.05）； the expression level of Beclin-1 protein and the ratio of LC3-Ⅱ/ LC3-Ⅰ in the cells in MPP⁺+mimics group and MPP⁺+FMF group were significantly higher than those in model group （P<0.01）； the expression level of Beclin-1 protein and the ratio of LC3-Ⅱ/ LC3-Ⅰ in the cells in MPP⁺+mimics+FMF group were significantly higher than those in MPP⁺+FMF group （P<0.01）. Conclusion FMF may promote the autophagy， alleviate the mitochondrial dysfunction， and attenuate the injury of the SH-SY5Y cells induced by MPP⁺， and its mechamism is related to upregulating the miR-145-3p expression.

Objective To discuss the change of reticulocyte hemoglobin （RET-He） level of the anemic patients with different degrees and different types of anemia， and to analyze its auxiliary diagnostic value in microcytic hypochromic anemia. Methods The clinical data of 369 anemic patients and 40 healthy physical examination individuals were collected. The patients were divided into mild anemia group （125 cases）， moderate anemia group （163 cases）， and severe anemia group （81 cases） based on the hemoglobin （Hb） level，and 40 healthy physical examination individuals were regarded as control group. The patients were also divided into macrocytic anemia group （91 cases）， normocytic anemia group （108 cases）， simple microcytic anemia group （23 cases）， and microcytic hypochromic anemia group （147 cases） based on the morphological types of anemia. The red blood cell count， Hb level， blood cell specific volume （HCT）， mean red blood cell volume （MCV）， mean red blood cell Hb level （MCH），mean red blood cell Hb concentration （MCHC）， and RET-He level of the subjects in various groups were detected；the receiver operating characteristic （ROC） curve was drawn to obtain the diagnostic cutoff value for the disease； the area under curve （AUC）， sensitivity， and specificity were calculated. Results Compared with control group， the RET-He levels of the patients in moderate and severe anemia groups were significantly decreased （P<0.01）； compared with mild anemia group， the RET-He levels of the patients in moderate and severe anemia groups were significantly decreased（P<0.01）. The RET-He levels of the patients in macrocytic anemia， normocytic anemia， simple microcytic anemia， and microcytic hypochromic anemia groups showed a decreasing trend （P<0.01）； compared with control group， the RET-He levels of the patients in simple microcytic anemia and microcytic hypochromic anemia groups were significantly decreased （P<0.01）； compared with mild macrocytic anemia group， the RET-He levels of the patients in mild normocytic anemia， mild simple microcytic anemia， and mild microcytic hypochromic anemia groups were significantly decreased （P<0.01）. Compared with mild normocytic anemia group， the RET-He levels of the patients in mild simple microcytic anemia group and mild microcytic hypochromic anemia group were significantly decreased （P<0.01）. Compared with moderate macrocytic anemia group， the RET-He levels of the patients in moderate normocytic anemia group， moderate microcytic anemia group， and moderate microcytic hypochromic anemia group were significantly decreased （P<0.05 or P<0.01）；compared with moderate normocytic anemia group， the RET-He level of the patients in moderate microcytic hypochromic anemia group was significantly decreased （P<0.01）； compared with severe macrocytic anemia group and severe normocytic anemia group， the RET-He level of the patients in severe microcytic hypochromic anemia group was significantly decreased （P<0.01）；compared with moderate simple microcytic anemia group， the RET-He level of the patients in severe simple microcytic anemia group was significantly increased （P<0.01）； compared with mild microcytic hypochromic anemia group， the RET-He levels of the patients in moderate microcytic hypochromic anemia group and severe microcytic hypochromic anemia group were significantly decreased （P<0.01）；compared with moderate microcytic hypochromic anemia group， the RET-He level of the patients in severe microcytic hypochromic anemia was significantly decreased （P<0.01）.The cutoff value of RET-He level for diagnosing microcytic hypochromic anemia was 27.25 pg， and the sensitivity was 91.2%， the specificity was 90.5%， and the AUC was 0.968［95% confidence interval （CI）：0.952—0.984］， suggesting that the RET-He level could make an auxiliary diagnosis for microcytic hypochromic anemia. Conclusion The RET-He level can assist in the diagnosis of anemia with different morphological types and degrees， and has certain clinical application value for the diagnosis of microcytic hypochromic anemia.