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

Organ structure destruction and functional decline leading to failure due to fibrosis pose severe threats to the human health and life. Macrophages are important immune cells present in various tissues and organs. Under the influence of numerous factors and pathways， such as transforming growth factor-β1（TGF-β1）， Toll-like receptor 4（TLR4）/nuclear factor-κB（NF-κB）， Janus kinase（JAK）/signal transducer and activator of transcription（STAT）， Notch signaling pathway， peroxisome proliferator-activated receptor（PPAR）， and cAMP response element-binding protein（CREB）， the macrophages may undergo polarization. In visceral organ fibrosis， the macrophage polarization signaling network， composed of single or multiple factors and pathways， is one of the crucial mechanisms regulating fibrosis. After polarization， the macrophages produce the chemokines and matrix metalloproteinase（MMP）， which are related to promoting fibrosis， leading to the occurrence and development of fibrosis. The current research conducted domestically and internationally mainly focuses on the roles of macrophages in infection， tumors， and fibrosis， with few summaries on the mechanisms of macrophage polarization in fibrosis. This review summarizes the pathways involved in macrophage polarization and the mechanisms of macrophage polarization in organ fibrosis， and provide the basis for the targeted therapy of fibrosis.

Objective To discuss the effect of hypoxia inducible factor-1α （HIF-1α）/ reactive oxygen species （ROS） on the apoptosis and invasion of the non-small cell lung cancer A549 cells，and to clarify the related mechanism. Methods The A549 cells were cultured under hypoxia condition for 12， 24， 48 and 72 h to construct the hypoxia cell model. The expression levels of HIF-1α mRNA in the cells at different treatment time were detected by real-time fluorescence quantitative PCR（RT-qPCR） method， and the proliferation abilities of the cells were detected by CCK-8 assay；the hypoxia cell model was verified， and the optimal induction time was determined.The A549 cells were divided into control group（21%O₂）， model group（hypoxia induction）， NAC group（20 mmol·L^－1 NAC）， NAC+YC-1 group（20 mmol·L^－1 NAC+100 μmol·L^－1 YC-1） and YC-1 （100 μmol·L^－1 YC-1）group. The expression levels of HIF-1α and FPR1 mRNA and proteins were detected by RT-qPCR and Western blotting methods； the ROS levels were detected by flow cytometry； the autophagosome structure in the cells was observed by transmission electron microscope； the expressions of microtubule-associated protein light china 3 Ⅱ（LC3-Ⅱ） in the cells in various groups were detected by immunofluorescence； the apoptotic rates of the cells in various groups were detected by flow cytometry； and the number of invasion cells was detected by Transwell chamber assay. Results With the prolongation of hypoxia induction time， the HIF-1α mRNA expression levels in the A549 cells and cell proliferation abilities were increased（P<0.05 or P<0.01）， and the optimal hypoxia induction time was 24 h. The results of RT-qPCR and Western blotting methods showed that compared with control group，the expression levels of HIF-1α and FPR1 mRNA and proteins in the cells in model group were significantly increased（P<0.01）； compared with model group，the expression levels of HIF-1α and FPR1 mRNA and proteins in the cells in NAC group， NAC+YC-1 group and YC-1 group were significantly decreased（P<0.01）.Compared with control group，the level of ROS in the cells in model group was significantly increased （P<0.01）； compared with model group，the ROS levels in the cells in NAC group， NAC+YC-1 group， and YC-1 group were significantly decreased（P<0.01）；compared with NAC group，the ROS level in the cells in NAC+YC-1 group was significantly decreased（P<0.01）.The transmission electron microscope results showed that the organelle structure of the cells in control group was intact and no autophagosome structure was seen； a large number of autophagosomes were seen in the cells in model group， obvious vacuoles were visible， and intracellular organelles were degraded； compared with model group， the number of autophagosomes in the cells in NAC group， NAC+YC-1 group and YC-1 group were decreased，among them the autophagosomes in the cells in NAC+YC-1 group were the least. The immunofluorescence assay results showed that the expression intensity of LC3-Ⅱ in the cells in model group was significantly increased compared with control group， and the expression intensities of LC3-Ⅱ in the cells in NAC group， NAC+YC-1 and YC-1 group were signficantly decreased compared with model group.Compared with control group， the apoptotic rate of the cells in model group was significantly decreased（P<0.01）； compared with model group， the apoptotic rates of the cells in NAC group， NAC+YC-1 group and YC-1 group were significantly increased（P<0.01）； compared with NAC group，the apoptotic rate of the cells in NAC+YC-1 group was increased（P<0.01）.The results of Transwell chamber assay showed that compared with control group the number of the invasion cells in model group was significantly increased（P<0.01）； compared with model group， the numbers of invasion cells in NAC group， NAC+YC-1 group and YC-1 group were significantly decreased （P<0.01）；compared with NAC group，the number of the in vasion cells in NAC+YC-1 group was decreased（P<0.01）. Conclusion Inhibition of HIF-1α/ROS under hypoxia condition can promote the apoptosis and inhibit cell invasion of cancer cells，and its mechanism may be realted to inhibiting the autophagy of lung cancer cells.

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 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 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 ameliorative effect of a novel antiepileptic drug Q808 on neuronal injury in temporal lobe epilepsy （TLE） rats， and to clarify its mechanism of action. Methods TLE rat model was prepared by intraperitoneal injection of the innovative antiepileptic drug candidate 6-（4-chlorophenoxy）- tetrazolo^{（5，1-a）} phthalazine （Q808）. Forty-five successfully modeled rats were randomly divided into model group， low dose of Q808 group， and high dose of Q808 group， and there were 15 rats in each group. The rats in low dose of Q808 group and high dose of Q808 group were gavaged with 20 and 80 mg·kg^-1 Q808，respectively， and the rats in model group were gavaged with an equal amount of 0.3% sodium carboxymethyl cellulose. Another 15 healthy SD rats were selected as control group. After 4 weeks of continuous gavage treatment， the morphology of the rats in varioius groups was observed； PONEMAH 6.X experimental animal telemetry platform was used to record the electroencephalogram of the rats in various groups； Golgi staining was used to observe the morphology of dendritic and dendritic spine density of hippocampal CA1 neurons of the rats in various groups； Western blotting method was used to detect the expression levels of synaptic plasticity-specific protein calcium/calmodulin-dependent protein kinaseⅡ （CaMKⅡ） in hippocampus tissue of the rats in various groups. Results The rats in control group showed normal activity without convulsions or other abnormal manifestations. The rats in model group， low dose of Q808 group， and high dose of Q808 group showed varying degrees of reduced activity， trembling and nodding， loss of balance， muscle rigidity and forelimb convulsions， gradually transforming into whole-body muscle rigidity and standing， followed by falling backwards， and there were no convulsions during the interictal period. Compared with control group， the total durations of epileptic seizures of the rats in model group， low dose of Q808 group， and high dose of Q808 group were significantly prolonged （P<0.01）. Compared with model group， the total durations of epileptic seizures in low dose of Q808 group and high dose of Q808 group were significantly shortened （P<0.01）. The hippocampal CA1 neurons of the rats in control group showed regular distribution of dendrites with dense and orderly dendritic networks. The hippocampal CA1 neurons of the rats in model group showed disordered arrangement of dendrites with massive dendritic entanglement， forming thicker nerve fiber bundles. Compared with model group， the dendritic networks of hippocampal CA1 neurons of the rats in low dose of Q808 group and high dose of Q808 group were partially recovered with relatively regular arrangement. Compared with control group， the dendritic spine density of hippocampal CA1 neurons of the rats in model group was significantly decreased （P<0.01）. Compared with model group， the dendritic spine densities of hippocampal CA1 neurons in low dose of Q808 group and high dose of Q808 group significantly increased （P<0.01）. Compared with control group， the expression levels of CaMKⅡ protein in hippocampus tissue of the rats in model group， low dose of Q808 group， and high dose of Q808 group were significantly decreased （P<0.01）. Compared with model group， the expression levels of CaMKⅡ protein in hippocampus tissue of the rats in low dose of Q808 group and high dose of Q808 group were significantly increased （P<0.01）. Conclusion The novel antiepileptic drug Q808 has an ameliorating effect on the TLE model rats；its mechanism may be related to Q808’s ability to reduce the dendritic lesions in hippocampal CA1 neurons and increase the expression level of synaptic plasticity-related protein CaMKⅡ protein.

Objective To discuss the expression of apoptosis-related gene CD44 in the thyroid cancer （THCA） tissue and its relationships with the clinicopathological characteristics of the patients and tumor infiltrating leukocytes（TILs）， and to provide new research directions for the diagnosis and treatment of THCA. Methods The expression spectrum of differentially expressed genes （DEGs） related to THCA apoptosis were obtained from The Cancer Genome Atlas （TCGA） Database. The up-regulated CD44 gene was selected and the expression level of CD44 mRNA in THCA tissue and parancancerous tissue were detected； receiver operating characteristic （ROC） curve was used to identify the THCA tissue and paracancerous tissue； Gene Ontology （GO）， Kyoto Encyclopedia of Genes and Genomes （KEGG）， and Gene Set Enrichment Analysis （GSEA） were used to analyze the function and pathway enrichment of DEGs； the protein-protein interaction （PPI） network was established by using STRING Database and visualized by Cytoscape software； the relationship between the expression of CD44 mRNA in THCA tissue and the infiltration abundance of TILs was analyzed by TIMER Database； the relationships between expression of CD44 protein and immune checkpoints were analyzed by R software. A total of 110 samples of THCA patients were selected， the expressions of CD44 protein in THCA tissue and paracancerous tissue were detected by the immunohistochemical SABC method. Results The TCGA Database analysis results showed that compared with paracancerous tissue，the expression level of CD44 mRNA in THCA tissue was increased（P<0.05）； the ROC curve analysis results showed that when the critical value was 6.855， the sensitivity， specificity， and accuracy of CD44 were 89.71%，75.13%， and 72.91%， respectively. The GO and KEGG analysis results showed that the DEGs were mainly enriched in the apoptosis-related pathways； the GSEA analysis results showed that the DEGs were associated with degranulation response of the neutrophils.The TIMER Database analysis results showed that the expression level of CD44 mRNA in THCA tissue was positively correlated with the infiltration abundance of B cells， macrophages， CD4+T lymphocytes， dendritic cells， and neutrophils （partial.cor>0，P<0.01），and was negatively correlated with tumor purity and the infiltration adundance of CD8+ T lymphocytes（partial.cor≤0，P<0.01）；the expression of CD44 protein in THCA tissue was positively correlated with 9 immune checkpoint genes （P<0.01）， and negatively correlated with 1 immune checkpoint gene （P<0.01）； the immunohistochemical SABC results showed that the positive expression rate of CD44 protein in THCA tissue was higher than that in paracancerous tissue （P<0.01）； the expression of CD44 protein in THCA tissue was correlated with the tumor diameter and lymph node metastasis （P<0.05），and was not correlated with gender， age， and extra glandular invasion of the patients（P>0.05）. Conclusion High expression of CD44 in THCA tissue may be related to progression and multiple immune responses of tumor.

Objective To discuss the clinical and pathological characteristics of IgA nephropathy （IgAN） in the children and adults， and to clarify their clinical significances. Methods The clinical and pathological data of the patients diagnosed with primary IgAN were collected. According to their ages， the patients were divided into children group （n=160） and adult group （n=240）. The ages，genders，incidences of hypertension，time from onset to renal biopsy，estimated glomerular filtration rates，initial onset manifestations，clinical types，Lee’s grades， MEST-C scores，immunofluorescence types and ultrastructures of glomeruli of the patients in two groups were compared. Results The ratio of male to female in children group was 1.5∶1 and it was 1.1∶1 in adults group； compared with adult group， the incidence of hypertension and the time from onset to renal biopsy of the patients in child group were decreased （P<0.05）， while the eGFR was increased（P<0.05）. Compared with adult group， the percentages of gross hematuria and edema of the patients in child group were increased（P<0.01）， and the percentages of abnormal urine test， foam urine， and other manifestations were decreased（P<0.01）.Compared with adult group， the percentages of the patients with isolated hematuria and nephrotic syndrome in child group were increased （P<0.01）， while the percentages of the patients with isolated proteinuria and chronic nephritis were decreased （P<0.01）；compared with adult group， the percentage of the patients with grade Ⅱ in the Lee’s grades in child group was increased（P<0.01） and the percentage of the patients with grade Ⅴ in the Lee’s grades was decreased （P<0.01）.The light microscope obervation results showed that there were only focal mesangial cells and mesangial matrix hyperplasia in grade Ⅱ renal tissue of the patients in child group， rarely accompanied by glomerulosclerosis. The renal tubular and interstitial lesions were not significant； there were significant proliferations of the mesangial cells and matrix in grade Ⅴ renal tissue of the patients in adult group， with more glomerulosclerosis， and relatively severe renal tubular and interstitial lesions. Compared with adult group， the percentages of the patients with M1 and E1 in the MEST-C scores of the patients in child group were increased （P<0.05 or P<0.01）， and the percentages of the patients with S1 and T1/T2 were decreased （P<0.01）.The urinary protein grade of the patients in child group was positively correlated with M（r_s=0.462），E（r_s=0.342），and C（r_s=0.250）scores （P<0.01）；the urinary protein grade of the patients in adult group was positively correlated with M（r_s=0.217），E（r_s=0.145），S（r_s=0.187），T（r_s=0.269），and C（r_s=0.256）scores （P<0.01）；compared with adult group， the IgA+IgG+IgM deposition of the patients in child group was increased（P<0.05）， deposition rate of C3 was increased（P<0.01）， and the deposition rate of fibrinogen （Fib） was increased（P<0.01）. Conclusion There are significant differences in the clinical and pathological characteristics between the children and the adults with primary IgAN， which should be treated differently in the clinical diagnosis， treatment， and research.

Objective To discuss the antioxidant capacities of ten edible plant exosomes-like nanoparticles （ELNs） from ginger， green pepper， garlic， mushroom， lemon， yam， grapes， tomatoes， broccoli，and onions in vitro and their protective effects on oxidative injury of the PC12 cells induced by hydrogen peroxide，and to provide the basis for the further study on the ELNs. Methods The PC12 cells were divided into control group， exosome group， hydrogen peroxide group，and hydrogen peroxide+exosomes group. Ten types of ELNs were separated and extracted by using high-speed differential centrifugation. The in vitro antioxidant capacities of these 10 types of ELNs were detected by using 1，1-diphenyl-2-picryl-hydrazyl（DPPH） free radical scavenging system，2，2'-azinobis-（3-ethylbenzthiazoline-6-sulphonate（ABTS） cationic radical system， and ferric ion total reduction antioxidant power（FRAD） system. The viabilities of PC12 cells in various groups were detected by MTT method. Results The DPPH radical scavenging abilities of ten ELNs，from high to low， were mushroom， onion， ginger， lemon， grape， tomato， green pepper， broccoli， yam， and garlic； the capacities of ten ELNs to scavenge ABTS， from high to low， were ginger， mushroom， onion， lemon， yam， grape， green pepper， garlic， tomato， and broccoli； the total antioxidant capacities of ten ELNs in FRAP， from high to low， were ginger， green pepper， onion， mushroom， lemon， broccoli， grape， yam， tomato，and garlic. The antioxidant capacities of five ELNs with stronger antioxidant capacities were increased with the increasing concentration of the ELNs in the DPPH radical scavenging system， ABTS cationic radical system and FRAP system. The half maximal inhibitory concentration（IC₅₀） values of onion were 73.15， 123.02， and 83.00 g·L^－1， the IC₅₀ values of ginger were 124.07， 91.24， and 91.24 g·L^－1， the IC₅₀ values of mushroom were 310.44， 518.04，and 237.10 g·L^－1， the IC₅₀ of green pepper were 969.06， 847.32，and 237.10 g·L^－1 and the IC₅₀ values of lemon were 1 718.94， 544.38，and 962.12 g·L^－1； compared with control group， the viability of the PC12 cells in hydrogen peroxide group was decreased by about 30%. Compared with hydrogen peroxide group， the viability of the PC12 cells in hydrogen peroxide+ELNs groups （green pepper， lemon， yam， and broccoli） were increased by 21.08%， 26.2%， 11.72%， and 15.15%. Conclusion The ELNs from ginger， mushroom， lemon， and onion show stronger antioxidant capacities in the DPPH radical scavenging system， ABTS cationic radical system and FRAP system. The ELNs from green pepper， lemon， yam， and broccoli have protective effects on the oxidative damage of the PC12 cells induced by hydrogen peroxide.

Early start denver model (ESDM) is a comprehensive early intervention approach for the children with autism spectrum disorder (ASD) between 12-month-old－36-month-old. The model is built upon the theoretical foundations of applied behavior analysis, denver model (DM), and pivotal response treatment, and it is one of the naturalistic developmental behavioral interventions. Compared with the other early intervention methods, ESDM is not limited by the environment of intervention; it encompasses all the areas of development during teaching practice and has been widely adopted for the early intervention of the children with ASD, and achieves the satisfactory therapeutic effect. The ESDM typically uses an intensive one-on-one intervention approach, but variabilities have emerged in its practical application, such as group ESDM(G-ESDM), parent-implemented ESDM (P-ESDM), and peer-mediated ESDM. In particular,G-ESDM and P-ESDM have provided the learning opportunities for more families, showing a broad application prospect. This study reviews the theoretical foundations, teaching models, and the effects of various intervention modalities of the ESDM in the treatment of ASD; combined with the domestic and international research findings,this study offers a reference for further studies on the mechanism of ESDM intervention for ASD.

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 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 screen the main characteristic variables affecting the incidence of cardiovascular and cerebrovascular diseases， and to construct the Bayesian network model of cardiovascular and cerebrovascular disease incidence risk based on the top 10 characteristic variables，and to provide the reference for predicting the risk of cardiovascular and cerebrovascular disease incidence. Methods From the UK Biobank Database， 315 896 participants and related variables were included. The feature selection was performed by categorical boosting （CatBoost） algorithm， and the participants were randomly divided into training set and test set in the ratio of 7∶3. A Bayesian network model was constructed based on the max-min hill-climbing （MMHC） algorithm. Results The prevalence of cardiovascular and cerebrovascular diseases in this study was 28.8%. The top 10 variables selected by the CatBoost algorithm were age， body mass index （BMI）， low-density lipoprotein cholesterol （LDL-C）， total cholesterol （TC）， the triglyceride-glucose （TyG） index， family history， apolipoprotein A/B ratio， high-density lipoprotein cholesterol （HDL-C）， smoking status， and gender. The area under the receiver operating characteristic （ROC） curve （AUC） for the CatBoost training set model was 0.770， and the model accuracy was 0.764； the AUC of validation set model was 0.759 and the model accuracy was 0.763. The clinical efficacy analysis results showed that the threshold range for the training set was 0.06-0.85 and the threshold range for the validation set was 0.09-0.81. The Bayesian network model analysis results indicated that age， gender， smoking status， family history， BMI， and apolipoprotein A/B ratio were directly related to the incidence of cardiovascular and cerebrovascular diseases and they were the significant risk factors. TyG index， HDL-C， LDL-C， and TC indirectly affect the risk of cardiovascular and cerebrovascular diseases through their impact on BMI and apolipoprotein A/B ratio. Conclusion Controlling BMI， apolipoprotein A/B ratio， and smoking behavior can reduce the incidence risk of cardiovascular and cerebrovascular diseases. The Bayesian network model can be used to predict the risk of cardiovascular and cerebrovascular disease incidence.

Objective To analyze the causal relationship between lifestyle-based factors and the occurrence and development of hepatobiliary malignancies by Mendelian randomization study method， and to provide the potential clinical evidence for the prevention and treatment of hepatobiliary malignancies. Methods The data from large-scale， independent genome-wide association studies （GWAS） were selected， and seven-step inclusion criteria for the instrumental variable screening were set up. The exposure lifestyles included the percentage of carbohydrate intake， percentage of fat intake， percentage of protein intake in the diet， coffee intake， weekly alcohol consumption times， leisure electronic screen exposure time， moderate to vigorous intensity physical activity （MVPA） during leisure time， sedentary behavior at work， age at first smoking， daily smoking quantity， current smoking status， and past smoking status， totaling 12 phenotypes. The primary analysis method used was the random effect model of the inverse variance weighted （IVW） method， and the heterogeneity was detected by Cochrane’s Q test and the horizontal pleiotropy was detected by MR-Egger intercept method. Results The current smoking status was significantly positively correlated with the increasing risk of extrahepatic cholangiocarcinoma （OR=1.607， 95% CI： 1.113-2.322， P=0.011）. Higher coffee intake was causally linked to a higher risk of liver cancer and intrahepatic cholangiocarcinoma （OR=1.000， 95% CI： 0.999-1.000， P=0.012）. In the physical activity， more MVPA was associated with the lower risk of liver cancer and intrahepatic cholangiocarcinoma （OR=0.998， 95% CI： 0.996-0.999， P=0.002）. The Cochrane’s Q test results showed that there was mild heterogeneity between MVPA and extrahepatic cholangiocarcinoma（Q=18.354，P=0.049） as well as the percentage of protein intake and intraphepatic cholangiocarainoma（Q=12.715，P=0.026）， and the MR-Egger intercept method results showed there was no horizontal pleiotropy. Conclusion There is a causal relationship between current smoking status and extrahepatic cholangiocarcinoma， and there is a causal relationship between more MVPA and the lower risk of liver cancer and intrahepatic cholangiocarcinoma. Education on smoking and physical activity for the patients may offer potential benefits for the prevention of hepatobiliary malignancies.

Chitooligosaccharides （COS）， as the degradation products of the natural polysaccharide chitosan （CS）， retain the good biocompatibility， non-toxicity， and biodegradability of CS. Additionally， due to the shortened sugar chains and reduced molecular weight， their water solubility and bioactivity are improved， making them more easily absorbed and utilized by organisms. In recent years， COS has received increasing attention. While there are numerous studies on the biological functions of CS and its applications in the biomedical field， the research on the functions and applications of COS is relatively limited. This study summarizes and analyzes the main biological functions of COS （anti-inflammatory， anti-tumor， antibacterial， and tissue regeneration promotion） and their possible mechanisms. It also discusses the research progress in the applications of COS in the biomedical field， in order to provide the theoretical basis and reference for the further research and broader application of COS in biomedicine.

Carpal tunnel syndrome （CTS） is one of the most common peripheral nerve entrapment disorders， the elevated pressure in the carpal tunnel， high-intensity activities and obesity are the main causes， and the patients with mild to moderate CTS are more prevalent. The main pathogenesis of CTS involves the increasing of carpal tunnel pressure and impaired local blood oxygen supply leading to reduced nerve conduction. Currently， the clinical treatment methods for mild to moderate CTS mainly include surgical and nonsurgical treatments. Nonsurgical treatment is the preferable choice for the patients with mild to moderate CTS. The western medical treatment primarily rely on oral medications， but their long-term use is limited due to the certain adverse effects； the local blockade and extracorporeal shock wave therapies show better efficacy for the patients with frequent activities and severe symptoms； the traditional Chinese medicine treatment also becomes a choice for some CTS patients due to their advantages of less pain， lower medical costs， and significant effectiveness. This study reviews the recent advancements in the pathogenesis and treatment of mild to moderate CTS， in order to design the personalized treatment methods for the mild to moderate CTS patients based on their specific conditions in clinical settings and provide the references for precise treatment of the mild to moderate CTS patients.

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 investigate the curative effect of osimertinib combined with savolitinib targeted therapy in the patients with non-small cell lung cancer（NSCLC） and central nervous system（CNS） metastasis with acquired resistance to epidermal growth factor receptor（EGFR）-tyrosine kinase inhibitor（TKI）， and to provide the reference for the targeted therapy for the disease. Methods The clinical data of one NSCLC complicated with CNS metastasis patient with non-frameshift deletion mutation in exon 19 of the EGFR gene complicated with MET amplification were collected， and the clinical characteristics， treatment methods， drug resistance mechanisms of targeted therapy， and treatment methods after drug resistance were analyzed combined with the literature review. Results The patient， female，47 years old， was diagnosed as lung adenocarcinoma in our hospital 3 years ago. The genetic test results showed that there was a non-frameshift deletion mutation in exon 19 of the EGFR gene， and the patient was given icotinib targeted therapy. After 18 months of treatment， the genetic test results showed the patient had the Exon20-T790M mutation， and the patient received oral osimertinib targeted therapy. Twelve months later， the disease progressed and CNS metastasis was found by the re-examination， and the genetic test showed the patient had the mesenchymal-epidermal transforming factor（MET） amplification，the patient received oral osimertinib combined with anlotinib targeted therapy. More than 2 months later， the re-examination results found that the disease had further progress， and the treatment plan was adjusted into osimertinib combined with savolitinib targeted therapy. After 1 month and 4 months， the re-examination results of chest CT and head MRI showed that the lung lesions and brain lesions were significantly smaller than before. Conclusion For the NSCLC brain metastases patients with EGFR-TKI resistance complicated with MET amplification after long-term targeted therapy， the combination of osimertinib and savolitinib targeted therapy can significantly control the progress of the lung and CNS disease in the short term.

The gastrointestinal motility disorder of the patients with Parkinson’s disease (PD) occurs in the early stages of this disease, even before the onset of motor symptoms. The gastrointestinal motility disorder is one type of gastrointestinal dysfunction, not only affect the absorption of medication, exacerbating the progression of PD, but also severely impact the quality of life of the patients. Therefore, it is essential to find new therapeutic targets to alleviate PD-induced gastrointestinal dysmotility in order to improve the progression of the disease and the quality of life of the patients. The gastrointestinal motility function is highly dependent on the health of the gut and central nervous regulating the gastrointestinal movements. A healthy gut is closely related to the integrity of the intestinal barrier, gut microbiota, neuroinflammation, and the normal function of enteric neurons responsible for the contraction and relaxation of the gastrointestinal tract. The gut function of the PD patients is compromised to some extent. This review summarizes the effects of the enteric nervous system, central nervous system, and gut microbiota on the development of gastrointestinal motility disorder of the PD patients;it also outlines the current therapeutic methods available and their limitations, with the aim of providing the new insights into the treatment of gastrointestinal motility disorder of the PD patients.

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.

Gag-Pol protein is one of the important structural proteins of human immunodeficiency virus （HIV）， comprising the basic scaffold proteins and functional enzymes required during the lifecycle of HIV. Currently，the inhibitors targeting different functional domains on Gag-Pol include capsid（CA） inhibitors，protease inhibitors， reverse transcriptase inhibitors， and integrase inhibitors. The CA inhibitors inhibit the maturation of CA or disrupt the assembly of CA，so as to affect the replication of HIV. The primary mechanism of protease inhibitors is to inhibit the protease from cleaving at the cleavage site CA-spacer peptide 1（SP1）. The reverse transcriptase inhibitors block the reverse transcription process of HIV by mimicking the reverse transcription substrates. The integrase inhibitors impact the activity of integrase by targeting the zinc-finger structure at the active center of integrase. This article summarizes the inhibitors targeting HIV Gag-Pol protein and their mechanisms， and reviews the approved dosages and usages of approved patent drugs，so as to provide the references for the future clinical combination therapies and the development of new inhibitors targeting HIV Gag-Pol protein.

Objective To screen the differentially expressed immune-related genes （DEIRGs） in in-stent restenosis （ISR）， and to analyze the immune cell infiltration in ISR， and to clarify the mechanism of occurrence and development of ISR. Methods The mRNA gene expression data of GSE46560 dataset samples were downloaded from the Gene Expression Omnibus （GEO），and divided into ISR group and non-ISR group. The “Limma” package in R software was used to identify the differentially expressed genes （DEGs） which were then intersected with immune-related genes （IRGs） to identify the DEIRGs in ISR； R software was used for Gene Ontology （GO） functional enrichment andalysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathway enrichment analysis on DEIRGs；the STRING database was used to construct the protein-protein interaction （PPI） network， which was visualized and analyzed for Hub genes by Cytoscape software； the receiver operating characteristic （ROC） curve of the Hub genes were plotted， and the area under the curve （AUC） was calculated and the diagnostic value was evaluated； CIBERSORT software was used to analyze the immune cell infiltration in ISR； Pearson correlation analysis was used to analyze the relationships between the immune cells and the relationships between the immune cells and key genes. Results A total of 331 DEGs were identified （P<0.05， | log₂FC| >1）， including 176 upregulated genes and 155 downregulated genes， and 38 DEIRGs were obstained. The GO functional enrichment analysis results showed that the DEIRGs were mainly enriched in biological processes （BP） such as defense response， immune response， and immune system； in cellular components （CC），the DEIRGs were located primarily in the extracellular region and cytoplasmic membrane； and in molecular functions （MF）， the DEIRGs were mainly involved in regulating signaling receptor binding and cytokine receptor activity.The KEGG signaling pathway enrichment analysis results indicated that the DEIRGs in ISR were primarily enriched in the phosphatidylinositol 3-kinase/protein kinase B （PI3K-AKT） and transforming growth factor-β （TGF-β） signaling pathways. In the PPI network， CD19 had the highest node among the top 10 Hub genes. Compared with non-ISR group， the expression level of the CD19 gene in the samples in ISR group was increased （P<0.05）. The AUC value in the ROC curve of CD19 gene expression was 0.92 （P<0.05）. The immune cell infiltration analysis results showed that compared with non-ISR group， the infiltration level of T lymphocyte follicular helper （Tfh） cells in the patients in ISR group were increased （P<0.05）， the infiltration levels of immature B lymphocytes， CD8+T lymphocytes， naive CD4+T lymphocytes， and M0 macrophages were increased， but the differences were not statistically significant （P>0.05）， while the infiltration levels of memory B lymphocytes， activated memory CD4+ T lymphocytes， regulatory T cells， resting natural killer （NK） cells， activated NK cells， monocytes， resting mast cells， and neutrophils were decreased， but the differences were not statistically significant （P>0.05）. There were positive correlations between Tfh cells and M0 macrophages and resting mast cells （r=0.88，P<0.05；r=0.68，P<0.05）， and there were negative correlations between Tfh cells and monocytes and neutrophils（r=-0.49，P<0.05；r=-0.42，P<0.05）. Conclusion CD19 may influence the occurrence and development of ISR by regulating the activation of the PI3K-AKT signaling pathway to affect the Tfh and B lymphocytes. CD19 can serve as a biomarker for the diagnosis of ISR.

Objective To discuss the expression level of macrophage scavenger receptor 1 （MSR1） in pan-cancer tissue，survival and immune characteristics of the patients by bioinformatics analysis， and to elucidate the value of MSR1 as a new biomarker for the tumor diagnosis， prognosis， and immunotherapy. Methods Clinical Bioinformatic Database and Sangerbox Database were used to analyze the expression levels of MSR1 mRNA in normal tissue and tumor tissue，the expression of MSR1 protein was analyzed by The Human Protein Atlas（HPA） Database，and univariate survival analysis and Kaplan-Meier analysis were used to evaluate the prognostic value of MSR1，and the expressions of MSR1 in various types of cells were analyzed with single-cell sequencing results from Tumor Immune Single-cell Hub （TISCH） Database；Tumor Immune Estimation Resource （TIMER2.0），Tumor Immune Syngeneic Mouse （TISMO）， Tumor Immune Dysfunction and Exclusion （TIDE）， and Gene Set Cancer Analysis （GSCA） Databases were used to analyze the correlations between the MSR1 expression level in pan-cancer tissue and immune cell infiltration， immune checkpoint gene expression， and immune treatment response. Results Compared with normal tissue， the expression levels of MSR1 mRNA in 17 kinds of tumor tissues including breast invasive carcinoma （BRCA），colon adenocarcinoma （COAD），esophageal carcinoma （ESCA）， glioblastoma multiforme （GBM）， head and neck squamous cell carcinoma （HNSC），kidney renal clear cell carcinoma （KIRC）， kidney renal papillary cell carcinoma （KIRP），brain low-grade glioma （LGG），liver hepatocellular carcinoma （LIHC），ovarian serous cystadenocarcinoma （OV），pancreatic adenocarcinoma（PAAD），prostate adenocarcinoma （PRAD），skin cutaneous melanoma （SKCM），stomach adenocarcinoma （STAD），testicular germ cell tumor （TGCT），thyroid carcinoma （THCA）， and uterine carcinosarcoma （UCS） were increased（P<0.01）；compared with normal tissue，the expression levels of MSR1 protein in breast cancer， endometrial cancer， liver cancer， ovarian cancer， skin melanoma， testis cancer， pancreatic cancer， and prostate cancer tissues were increased in varying degrees. The high expressions of MSR1 in bladder urothelial carcinoma ［BLCA， hazard ratio（HR）=1.01， P=0.047， 95%CI（1.00，1.03）］， LGG ［HR=1.03， P<0.001， 95%CI（1.02，1.04）］， LIHC ［HR=1.04， P=0.007， 95%CI（1.01，1.07）］， OV ［HR=1.02，P=0.028， 95%CI（1.00，1.03）］， STAD ［HR=1.02， P=0.016， 95%CI（1.00，1.04）］， THCA［HR=1.06，P=0.006， 95%CI（1.02，1.11）］ and uveal melanoma ［UVM， HR=1.18，P=0.044， 95%CI（1.00，1.40）］ were correlated with the poor overall survival（OS） of the patients； the high expression of MSR1 in LGG ［HR=1.03，P<0.001， 95%CI（1.02，1.04）］，the uterine corpus endometrial carcinoma （UCEC］， ［HR=1.05， P=0.038， 95%CI（1.00，1.10）］， and UVM ［HR=1.2，P=0.036， 95%CI（1.01，1.41）］ was correlated with poor disease-specific surival（DSS）.The single-cell sequencing results indicated that MSR1 was mainly expressed in the dendritic cell（DC） and monocyte-macrophage. The MSR1 expression was positive correlated with various kinds of immune cell infiltrations（P<0.05），including CD8+ T lymphocytes， natural killing（NK） cells， regulatory T lymphocytes， and cancer associated fibroblasts（CAF）. There were significant positive correlations between MSR1 and classical immune checkpoint gene and immunotherapy response marker （P<0.05）. Conclusion MSR1 is highly expressed in lots of tumor tissues and is closely associated with the poor prognosis and immune factors of lots of tumor patients. MSR1 may be a diagnostic tumor marker and a predictor of tumor prognosis and immunotherapy efficacy， and has the potential to be a new therapeutic target.

Objective To discuss the relationship between microdeletions of 15 sequence tagged sites （STS） in azoospermia factor （AZF） region of the Y chromosome and male infertility （MI）， and to provide the evidence for the intervention of hereditary MI. Methods A total of 2 586 suspected MI patients were selected， and they were divided into four groups according to their ages，≤20 years old group（14 cases）， 21—30 years old group（988 cases）， 31—40 years old group（1 318 cases），and ≥41 years old group（266 cases）. The polymerase chain reaction （PCR） method was used to detect the 15 STS sequence fragments in the Y chromosome AZF region and the abnormal results were screened out. The microdeletion status of the Y chromosome was compared among the MI patients in various groups. Results Among 2 586 samples， 207 cases of Y chromosome abnormalities were found， accounting for 8.00% （207/2 586） of the total samples. The Y chromosome abnormalit rates of the somples in≤20 years old， 21—30 years old，31—40 years old， and ≥41 years old groups were 7.14% （1/14）， 8.10% （80/988），8.04% （106/1 318）， and 7.52% （20/266）， respectively； there were significant differences in the detetion rates of base sites and extension sites of the patients between various groups （χ²=10.836，P=0.013），and the deletion rate of base sites and extension sites of the patients in 21—30 years old group was higher than that in 31—40 years old group （P<0.05）. In the overall tested samples， 52 cases of base site fragment deletion were found， the abnormality rate was 2.01%， and there were significant differences in the abnormality rates of the patients between various groups （χ²=9.658， P=0.022）. The deletion rate of the AZFc segment accounted for 1.39% of all tested individuals， and the deletion rates of the patients in 21—30 years old group and 31—40 years old group were significantly higher than that in ≥41 years old group （P<0.05）.There were significant differences in the overall deletion rates of the patients between 21—30 years old and 31—40 years old groups（χ²=3.612，P=0.040）. There were no statistically significant differences in the deletion rates of the sY127， sY134 combined with sY105， sY121， sY1192， sY153， and sY160 sites of the patients between various groups （P>0.05），and there were no significant differences in the deletion rates of the sY254， sY255 combined with sY105， sY121， sY1192， sY153， and sY160 sites of the patients between various groups （P>0.05）. Conclusion The main cause of Y chromosome abnormalities of the males in reproductive age group in Northeast of Guangxi Zhuang Autonomous Region is microdeletion at the sY1192 and sY153 sites， and the sY1192 site microdeletion is the most prevalent. The detection rate of mutation at this site is increased with the increasing of age.

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 discuss the improved methods for the isolation and culture of primary chondrocytes from the neonatal rats，and to establish an efficient and economical in vitro chondrocyte culture system. Methods The primary chondrocytes were isolated from the joints of neonatal rats and divided into overnight digestion （OD） group and rapid digestion （RD） group for separation. The chondrocytes in OD group were digested overnight by typeⅡ collagenase， while the chondrocytes in RD group were separated by the combination of pre-digestion with physical and chemical digestion methods. The chondrocytes were cultured in modified media containing 0% （blank group 1）， 1%， 2%， 4%， and 10% fetal bovine serum （FBS）， 0 （blank group 2）， 0.1， 0.2， 0.4， 0.8， 1.0， and 2.0 g·L^-1 vitamin C（VC）， and 0 （blank group 3）， 0.5， 1.0， 2.0， 4.0， 8.0， 10.0 μg·L^-1 poly（lactic-co-glycolic acid） （PLGA） nanoparticles. The media containing different concentrations of FBS， VC， and PLGA were mixed with Dulbecco’s modified Eagle’s medium/nutrient mixture F-12（DMEM/F12）， and were divided into related groups based on the concentrations of ingredients. Cell counter was used to count the chondrocytes in various groups and the survival rates and diameters of the chondrocytes in various groups were detected； Toluidine blue staining was used to detect the morphology of the chondrocytes in various groups； CCK-8 method was used to detect the proliferative activities of the chondrocytes in various groups； cell adhesion assay was used to detect the adhesion rates of the chondrocytes in various groups； Hoechst/propidium iodide（PI） staining was used to detect the apoptosis of the chondrocytes in various groups； MTT assay was used to detect the proliferation activities of the chondrocytes in various groups after treated with modified media.The cells were divided into DMEM/F12+10%FBS group， DMEM/F12+1%FBS group， and DMEM/F12+1% FBS+0.4 g·L^-1 VC+1 μg·L^-1 PLGA group. Real-time fluorescence quantitative PCR（RT-qPCR） method was used to detect the expression levels of sex-determining region Y-box 9 （SOX9）， collagen type Ⅱ alpha 1 chain （Col2A1）， collagen type Ⅹ alpha 1 chain （Col10A1）， and matrix metallopeptidase 13 （MMP13） mRNAs in the chondrocytes in various groups after treated with modified media；immunofluorescence staining was used to detect the expressions of type Ⅱ collagen （COLⅡ） and SOX9 in the chondrocytes in various groups after treated with modified media. Results The survival rate of primary chondrocytes in OD group was lower than that in RD group， and the average cell diameter was larger than that in RD group. The primary chondrocytes in OD group were larger and spindle-shaped， and most cells exhibited pseudopodia； in RD group， the primary chondrocytes were smaller， mostly rhomboid in shape， with only a portion of the cells showing pseudopodia. The Toluidine blue staining results showed significant coloration in both groups， but the digestion time of the chondrocytes in RD group was shorter， and compared with OD group， the actual culture time of the chondrocytes was reduced by 9-13 h， and more immature morphology of the primary chondrocytes were observed. The proliferation activity of the primary chondrocytes in OD group was slow at 24 h of culture but increased at 48 h of culture， and the proliferation activity of the primary chondrocytes was significantly higher at 48 h of culture compared with 12 h of culture （P<0.01）. Compared with 12 h of culture，the proliferation rates of the primary chondrocytes in RD group were increased at 24 and 48 h of culture （P<0.01）. At 24 and 48 h of culture， compared with OD group， the proliferation rates of the primary chondrocytes in RD group were increased （P<0.05）. The number of apoptotic chondrocytes in RD group was lower than that in OD group， and no necrotic chondrocytes were observed in either group. The proliferation activities of chondrocytes of the rats were increased with the rising of FBS concentration in the culture medium. Compared with blank group 1， the proliferation activities of chondrocytes of the rats after treated with culture mediums containing 1%， 2%， 4%， and 10% FBS were significantly increased （P<0.05）. Compared with blank group 2， the proliferative activities of chondrocytes of the rats after treated with culture mediums containing 0.2-1.0 g·L^-1 VC were significantly increased （P<0.05）， and the highest proliferation activity was found when the concentration of VC was 0.4 g·L^-1 （P<0.01）. Compared with blank group 3， the proliferation activities of chondrocytes of the rats after treated with culture mediums containing 1-4 μg·L^-1 PLGA were significantly increased （P<0.05）， and the highest proliferation activity was found after treated with culture medium containing 1 μg·L^-1 PLGA （P<0.05）. Compared with DMEM/F12+10%FBS group， the expression levels of SOX9 mRNA and Col2A1 mRNA in the chondrocytes in DMEM/F12+1%FBS group were significantly increased （P<0.05 or P<0.01）. Compared with DMEM/F12+10%FBS group， the expression levels of SOX9 mRNA and Col2A1 mRNA in the chondrocytes in DMEM/F12+1%FBS+0.4 g·L^-1 VC+1 μg·L^-1 PLGA group were significantly increased （P<0.01）. The immunofluorescence staining results showed that the green fluorescence signal of COLⅡ and the red fluorescence signal of SOX9 were observed in some chondrocytes in DMEM/F12+10%FBS group under fluorescence microscope， and the fluorescence intensity was weak. In DMEM/F12+1%FBS group， most chondrocytes exhibited COLⅡ green fluorescence signal and SOX9 red fluorescence signal， and the fluorescence intensity was significantly stronger than that in DMEM/F12+10% FBS group. In DMEM/F12+1% FBS+0.4 g·L^-1 VC+1 μg·L^-1 PLGA group， the COLⅡ green fluorescence signal and SOX9 red fluorescence signal were found in all the chondrocytes， and the fluorescence intensity was significantly higher than those in DMEM/F12+10%FBS and DMEM/F12+1%FBS groups. The expression levels of COLⅡ and SOX9 proteins in the chondrocytes in DMEM/F12+1%FBS group were significantly higher than those in DMEM/F12+10%FBS group， and the expression levels of COLⅡ and SOX9 proteins in the chondrocytes in DMEM/F12+1%FBS+0.4 g·L^-1 VC+ 1 μg·L^-1 PLGA group were significantly higher than those in DMEM/F12+10%FBS group. Conclusion The improved methods for the isolation and culture of primary chondrocytes of the rats can overcome the shortcomings of traditional methods， shorten the isolation time of primary chondrocytes， and improve the quality of in vitro culture of primary chondrocytes.

Objective To detect the levels and existing forms of vitamin D （VD） in peripheral blood of the children with development language disorder（DLD） comorbid with attention deficit hyperactivity disorder（ADHD），to clarify the role of VD in the occurrence of DLD comorbid with ADHD， and to provide the new method for its treatment. Methods A total of 90 children with DLD comorbid with ADHD were regarded as observation group， and fifty physically and mentally healthy children in the Health Examination Center in our hospital during the same period were selected as control group. The DLD and ADHD were diagnosed according to the relevant chapters of the International Classification of Diseases， 11th Revision （ICD-11） and the American Diagnostic and Statistical Manual of Mental Disorders， Fifth Edition （DSM-5）.Griffiths Mental Development Scales （GMDS） was used to detect the language ability and the Developmental Quotient（DQ） was caculated；ADHD was classified according to the Snoopy Rating Scale Ⅳ （SNAP-4） Parent and Teacher Questionnaire；Conners Parent Symptom Questionnaire （PSQ） was used for ADHD scoring；the levels of serum VD of the subjects in two groups were detected by liquid chromatography-mass spectrometry. The results of GMDS and PSQ of all subjects and SNAP-4 of the subjects in observation group were collected.When DQ was less than （100－1）×standard deviation， the language ability of the subjects was lower than that of the contemporary；the children’s behavioral problems were divided into six factors according to the PSQ，such as learning problem， psychosomatic problem， anxiety， hyperactivity-impulsivity， conduct problem， and hyperactivity index； when the scores of six factors were all greater than 2 points， the related problems could be diagnosed；according to SNAP-4， ADHD was divided into predominantly inattentive ADHD（ADHD-I），predominantly-hyperactivity/impulsivity（ADHD-HI） and combined ADHD（ADHD-C） groups. Results In observation group， 71.1% were male and 28.9% were female. The levels of serum VD3 and VD of the chinldren in observation group were significantly lower than those in control group （P<0.01）.The levels of serum VD and VD3 of all three types（DLD-ADHD-C，DLD-ADHD-HI，and DLD-ADHD-I） of the subjects in observation group were significantly lower than those in control group （P<0.01）.The DQ values of the subjects in control group were positively correlated with the levels of serum VD and VD3（r=0.512，P<0.01；r=0.529，P<0.01）；the DQ values of the subjects in observation group was positively correlated with the levels of serum VD and VD3（r=0.299， P<0.01； r=0.279， P<0.01），and was negatively correlated with the level of VD2（r=－0.122， P<0.01）.There were significantly differences in the scores of learning problem， psychosomatic problem， conduct problem，anxiety， hyperactivity/impulsivity and hyperactivity index among three types （DLD-ADHD-C，DLD-ADHD-HI，and DLD-ADHD-I） of the subjects in observation group（P<0.01）.The level of serum VD3 was negatively correlated with the scores of the above indexes of the chlildren with DLD-ADHD-C type in observation group（r=－0.438，r=－0.357，r=－0.422，r=－0.465，r=－0.583， r=－0.593，P<0.01），and was positively correlated with the conduct problem of the children with DLD-ADHD-HI type in observation group（r=0.522，P<0.01），the level of VD3 was negatively correlated with hyperactivity-impulsivity and hyperactivity index（r=－0.455，P<0.05； r=－0.424，P<0.01），was negatively correlated psychosomatic problems and hyperactivity-impulsivity（r=－0.468，r=－0.496，P<0.05）， and was positively correlated with hyperactivity index（r=0.694，P<0.01）. The level of VD was negatively correlated with the scores of all six factors mentioned above of the children with DLD-ADHD-C type in observation group（r=－0.444，r=－0.498，r=－0.450，r=－0.501，r=－0.594， r=－0.522，P<0.01），and was negatively correlated with psychosomatic problem， conduct problem，hyperactivity/impulsivity and hyperactivity index of the children with DLD-ADHD-HI type in observation group（r=－0.355，r=－0.578，r=－0.509， r=－0.422，P<0.05or P<0.01）；the level of VD was negatively correlated with psychosomatic problem and hyperactivity/impulsivity（r=－0.485，r=－0.497，P<0.05）， and was positively correlated with the hyperactivity index （r=0.682，P<0.01）. Conclusion VD3 deficiency may be one of the causes of DLD comorbid with ADHD.VD3 supplementation may have a positive effect in the prevention and treatment of the children with DLD comorbid with ADHD.