BACKGROUND:Macrophage migration inhibitory factor(MIF)is a pleiotropic cytokine,which is secreted in different types of stem cells and can regulate the proliferation,differentiation and migration of various types of stem cells.Our previous research has confirmed that human embryonic stem cells secrete MIF and that its concentration in the culture medium is relatively stable.However,whether MIF is involved in the survival,proliferation and differentiation of human embryonic stem cells remains unclear. OBJECTIVE:To investigate the effects of MIF on survival,proliferation,and differentiation of human embryonic stem cells. METHODS:(1)Human embryonic stem cells H9 were cultured.The growth curve of cells was detected and plotted by CCK-8 assay.Enzyme-linked immunosorbent assay was used to determine the level of MIF in the medium.(2)To determine the effects of exogenous MIF on the survival and proliferation of human embryonic stem cells,different groups were established:the control group,which was cultured in stem cell medium without any modifications;the exogenous MIF group,which was treated with different concentrations(30,100,300 ng/mL)of MIF in the stem cell medium;the MIF inhibitor ISO-1 group,which was treated with different concentrations(2,7,21 μmol/L)of ISO-1 in the stem cell medium;and the MIF+ISO-1 group,which was treated with different concentrations of ISO-1 along with 100 ng/mL of MIF.Cell viability was assessed using the CCK-8 assay.(3)To further elucidate the effect of MIF gene on survival and proliferation of human embryonic stem cell,the MIF knockout H9 cell line was constructed by CRISPR-Cas 9 technology to observe the lineage establishment.(4)To determine the effect of high concentrations of MIF on human embryonic stem cell differentiation,100 ng/mL MIF and 100 ng/mL of CXCR4 neutralizing antibody were separately added to the normal stem cell culture medium.The expression levels of self-renewal factors(KLF4,c-MYC,NANOG,OCT4,and SOX2)and differentiation transcription factors(FOXA2,OTX2)were measured using real-time quantitative polymerase chain reaction,immunofluorescence staining,and western blot analysis. RESULTS AND CONCLUSION:(1)The logarithmic growth phase of H9 cells was between 3-6 days.Under normal growth conditions,human embryonic stem cells secreted MIF at a concentration of approximately 20 ng/mL,independent of cell quantity.(2)Compared to the control group,the addition of different concentrations of MIF had no effect on the proliferation of human embryonic stem cells(P>0.05).ISO-1 significantly inhibited the proliferation of human embryonic stem cells,with a stronger inhibition observed at higher concentrations of ISO-1(P<0.05).The addition of MIF in the presence of ISO-1 reduced the inhibitory effect of ISO-1(P<0.05).(3)Real-time quantitative polymerase chain reaction showed that knocking out 50%of the MIF gene resulted in a significant decrease in the growth vitality of human embryonic stem cells and failure to establish cell lines.(4)Adding 100 ng/mL exogenous MIF to the culture medium resulted in a decrease in the mRNA,protein,and fluorescence expression levels of the self-renewal transcription factor KLF4,while the mRNA,protein,and fluorescence expression levels of the differentiation factor FOXA2 increased.(5)When 100 ng/mL CXCR4 neutralizing antibody was added to the culture medium,the mRNA and protein expression levels of KLF4 increased,while the mRNA and protein expression levels of FOXA2 decreased,contrary to the expression trend observed in the MIF group.In conclusion,the endogenous secretion of MIF by human embryonic stem cells is essential for their survival.The addition of MIF to the culture medium does not promote the proliferation of human embryonic stem cells.However,it can lead to a decrease in the expression of the self-renewal factor KLF4 and an increase in the expression of the transcription factor FOXA2.This provides a clue for further investigation into the effects and mechanisms of MIF on the differentiation of human embryonic stem cells.The MIF-CXCR4 axis plays a regulatory role in this process.

ObjectiveTo explore the role of cucurbitacin B （CuB） in inducing ferroptosis in 4T1 cells and its mechanism. MethodsThe effects of CuB（0.2， 0.4， 0.8 μmol·L^-1）on the proliferation ability of 4T1 cells in vitro were detected using the methyl thiazolyl tetrazolium （MTT） assay. The clonogenic ability of 4T1 cells was detected by the plate cloning assay， and the levels of lactate dehydrogenase （LDH） in 4T1 cells were detected by the use of a kit. The mitochondrial membrane potential and reactive oxygen species （ROS） levels in 4T1 cells were detected by flow cytometry， and the mitochondrial ultrastructure of 4T1 cells was observed by transmission electron microscopy. The western blot was used to detect the expression of ferroptosis-related protein p53 in 4T1 cells， solute carrier family 7 member 11 （SCL7A11）， glutathione peroxidase 4 （GPX4）， long-chain acyl-CoA synthetase 4 （ACSL4）， transferrin receptor protein 1 （TFR1）， and ferritin heavy chain 1 （FTH1）. ResultsCompared with that in the blank group， the survival rate of 4T1 cells in CuB groups was significantly decreased （P<0.05）， and the number of cell clones in CuB groups was significantly reduced （P<0.01）. In addition， compared with that in the blank group， the leakage of LDH in cells in CuB groups was significantly increased （P<0.01）， and the mitochondrial membrane potential of cells in CuB groups decreased significantly （P<0.01）. Cellular ROS levels were significantly elevated in CuB groups （P<0.01）. The mitochondria of cells in CuB groups were obviously wrinkled， and the mitochondrial cristae were reduced or even disappeared. Compared with that in the blank group， the protein expression of p53， ACSL4， and TFR1 were significantly up-regulated in CuB groups （P<0.05）， and that of SLC7A11， GPX4， and FTH1 were significantly down-regulated （P<0.05）. ConclusionCuB may inhibit SLC7A11 and GPX4 expression by up-regulating the expression of p53， which in turn regulates the p53/SLC7A11/GPX4 signaling pathway axis and accelerates the generation of lipid peroxidation substrate by up-regulating the expression of ACSL4. It up-regulates TFR1 expression to promote cellular uptake of Fe³⁺ and down-regulates the expression of FTH1 to reduce the ability of iron storage， resulting in an elevated free Fe²⁺ level. It catalyzes the Fenton reaction， generates excess ROS， imbalances the antioxidant system and iron metabolism， and then induces ferroptosis in 4T1 cells.

ObjectiveTo establish a rapid analytical method for identifying the differential components in Poria cocos medicinal materials based on ultra performance liquid chromatography-quadrupole-electrostatic field orbital trap high-resolution mass spectrometry（UPLC-Q-Orbitrap-MS）， combined with mass defect filtering（MDF） and molecular network integration techniques. MethodsUPLC-Q-Orbitrap-MS was used for MS data acquisition and identification of P. cocos medicinal materials， with the help of MDF for the study of cleavage behavior and structural identification of triterpenoids. According to the similarity of MS/MS fragmentation patterns of each component， global natural product social molecular network（GNPS） was established， and Cytoscape 3.6.1 was used to screen molecular clusters with similar structures and the the structure of main compound classes were identified and confirmed. Multivariate statistical analyses such as principal component analysis（PCA） and orthogonal partial least squares-discriminant analysis（OPLS-DA） were used to screen the differential components of the five P. cocos medicinal materials with the variable importance in the projection（VIP） value>1 and P<0.05 as the criteria. ResultsA total of 66 compounds were identified by database comparison， 8 compounds were newly identified by MDF， 28 compounds were newly identified by GNPS， and a total of 102 chemical compounds were identified， including 43 triterpenoids， 16 saccharides， 26 amino acids and peptides， 3 nucleosides， and 14 other compounds. Triterpenoids were predominant in Poriae Cutis and wild Fushen， amino acids and peptides were the most abundant in Poria and cultivated Fushen， carbohydrates were the most abundant in Poriae Cutis. Type Ⅰ and Ⅱ triterpenoids had higher amounts in Poria and cultivated Fushen， type Ⅲ triterpenoids were more abundant in Poriae Cutis， all four types of triterpenoids were higher in Fushenmu， and type Ⅰ， Ⅱ， and Ⅳ triterpenoids were higher in wild Fushen. A total of 12 common differential chemical constituents were screened， including serine， guanosine， gallic acid， 2-octenal， maltotriose， trametenolic acid， dehydroeburicoic acid， dehydrotrametenolic acid， poricoic acid A， poricoic acid B， poricoic acid E and G， but the relative contents of them varied significantly among different medicinal materials. ConclusionAmong the five P. cocos medicinal materials， the types of constituents are generally similar， but their relative contents differed significantly among these medicinal materials， especially in the distribution of triterpenoids. The integration of UPLC-Q-Orbitrap-MS， MDF and GNPS can provide a reference for the rapid qualitative analysis of other Chinese medicines.

ObjectiveTo explore the role of cucurbitacin B （CuB） in inducing ferroptosis in 4T1 cells and its mechanism. MethodsThe effects of CuB（0.2， 0.4， 0.8 μmol·L^-1）on the proliferation ability of 4T1 cells in vitro were detected using the methyl thiazolyl tetrazolium （MTT） assay. The clonogenic ability of 4T1 cells was detected by the plate cloning assay， and the levels of lactate dehydrogenase （LDH） in 4T1 cells were detected by the use of a kit. The mitochondrial membrane potential and reactive oxygen species （ROS） levels in 4T1 cells were detected by flow cytometry， and the mitochondrial ultrastructure of 4T1 cells was observed by transmission electron microscopy. The western blot was used to detect the expression of ferroptosis-related protein p53 in 4T1 cells， solute carrier family 7 member 11 （SCL7A11）， glutathione peroxidase 4 （GPX4）， long-chain acyl-CoA synthetase 4 （ACSL4）， transferrin receptor protein 1 （TFR1）， and ferritin heavy chain 1 （FTH1）. ResultsCompared with that in the blank group， the survival rate of 4T1 cells in CuB groups was significantly decreased （P<0.05）， and the number of cell clones in CuB groups was significantly reduced （P<0.01）. In addition， compared with that in the blank group， the leakage of LDH in cells in CuB groups was significantly increased （P<0.01）， and the mitochondrial membrane potential of cells in CuB groups decreased significantly （P<0.01）. Cellular ROS levels were significantly elevated in CuB groups （P<0.01）. The mitochondria of cells in CuB groups were obviously wrinkled， and the mitochondrial cristae were reduced or even disappeared. Compared with that in the blank group， the protein expression of p53， ACSL4， and TFR1 were significantly up-regulated in CuB groups （P<0.05）， and that of SLC7A11， GPX4， and FTH1 were significantly down-regulated （P<0.05）. ConclusionCuB may inhibit SLC7A11 and GPX4 expression by up-regulating the expression of p53， which in turn regulates the p53/SLC7A11/GPX4 signaling pathway axis and accelerates the generation of lipid peroxidation substrate by up-regulating the expression of ACSL4. It up-regulates TFR1 expression to promote cellular uptake of Fe³⁺ and down-regulates the expression of FTH1 to reduce the ability of iron storage， resulting in an elevated free Fe²⁺ level. It catalyzes the Fenton reaction， generates excess ROS， imbalances the antioxidant system and iron metabolism， and then induces ferroptosis in 4T1 cells.

ObjectiveTo establish a rapid analytical method for identifying the differential components in Poria cocos medicinal materials based on ultra performance liquid chromatography-quadrupole-electrostatic field orbital trap high-resolution mass spectrometry（UPLC-Q-Orbitrap-MS）， combined with mass defect filtering（MDF） and molecular network integration techniques. MethodsUPLC-Q-Orbitrap-MS was used for MS data acquisition and identification of P. cocos medicinal materials， with the help of MDF for the study of cleavage behavior and structural identification of triterpenoids. According to the similarity of MS/MS fragmentation patterns of each component， global natural product social molecular network（GNPS） was established， and Cytoscape 3.6.1 was used to screen molecular clusters with similar structures and the the structure of main compound classes were identified and confirmed. Multivariate statistical analyses such as principal component analysis（PCA） and orthogonal partial least squares-discriminant analysis（OPLS-DA） were used to screen the differential components of the five P. cocos medicinal materials with the variable importance in the projection（VIP） value>1 and P<0.05 as the criteria. ResultsA total of 66 compounds were identified by database comparison， 8 compounds were newly identified by MDF， 28 compounds were newly identified by GNPS， and a total of 102 chemical compounds were identified， including 43 triterpenoids， 16 saccharides， 26 amino acids and peptides， 3 nucleosides， and 14 other compounds. Triterpenoids were predominant in Poriae Cutis and wild Fushen， amino acids and peptides were the most abundant in Poria and cultivated Fushen， carbohydrates were the most abundant in Poriae Cutis. Type Ⅰ and Ⅱ triterpenoids had higher amounts in Poria and cultivated Fushen， type Ⅲ triterpenoids were more abundant in Poriae Cutis， all four types of triterpenoids were higher in Fushenmu， and type Ⅰ， Ⅱ， and Ⅳ triterpenoids were higher in wild Fushen. A total of 12 common differential chemical constituents were screened， including serine， guanosine， gallic acid， 2-octenal， maltotriose， trametenolic acid， dehydroeburicoic acid， dehydrotrametenolic acid， poricoic acid A， poricoic acid B， poricoic acid E and G， but the relative contents of them varied significantly among different medicinal materials. ConclusionAmong the five P. cocos medicinal materials， the types of constituents are generally similar， but their relative contents differed significantly among these medicinal materials， especially in the distribution of triterpenoids. The integration of UPLC-Q-Orbitrap-MS， MDF and GNPS can provide a reference for the rapid qualitative analysis of other Chinese medicines.

Hearing loss has become increasingly prevalent and causes considerable disability, thus gravely burdening the global economy. Irreversible loss of hair cells is a main cause of sensorineural hearing loss, and currently, the only relatively effective clinical treatments are limited to digital hearing equipment like cochlear implants and hearing aids, but these are of limited benefit in patients. It is therefore urgent to understand the mechanisms of damage repair in order to develop new neuroprotective strategies. At present, how to promote the regeneration of functional hair cells is a key scientific question in the field of hearing research. Multiple signaling pathways and transcriptional factors trigger the activation of hair cell progenitors and ensure the maturation of newborn hair cells, and in this article, we first review the principal mechanisms underlying hair cell reproduction. We then further discuss therapeutic strategies involving the co-regulation of multiple signaling pathways in order to induce effective functional hair cell regeneration after degeneration, and we summarize current achievements in hair cell regeneration. Lastly, we discuss potential future approaches, such as small molecule drugs and gene therapy, which might be applied for regenerating functional hair cells in the clinic.
Infant, Newborn ; Humans ; Hair Cells, Auditory, Inner/physiology* ; Ear, Inner/physiology* ; Hair Cells, Auditory/physiology* ; Regeneration/genetics* ; Stem Cells

Objective:To understand the health status,influencing factors and spatial distribution of the Chinese floating population and to evaluate the health equity of the floating population.Methods:All the data were collected from the 2017 Migrant Population Dynamic Monitoring Survey in China,binary Logistic regression was used to analyze the factors that might affect the health of the floating population,and the concentration index method was used to evaluate the health equity of the floating population.Spa-tial autocorrelation analyses the spatial aggregation of health status and health equity.Results:The un-healthy rate of the floating population in China was 2.71％.Age and gender show a statistically signifi-cant impact on self-rated health;that is,as age increases,the self-rated health of the migrant population gradually deteriorates,and women are more likely to think that they are unhealthy.Fairness analysis shows that the concentration index of the floating population is 0.021 7,the urban household registration floating population is 0.021 6,and the rural household registration floating population is 0.021 9.It is shown that the fairness of the health status of the floating population is biased towards the high-income class,and the rural household registration floating population's health unfairness is greater than that of the urban household registration migration population.Moreover,Moran's i=0.211 for self-rated health and Moran's i=0.291 for the unhealthy rate indicate that self-rated health has a spatial aggregation trend.Moran's i=0.136 showed the characteristics of spatial clustering,and the two-week prevalence fairness of the floating population was mainly in the northern and southeastern coastal areas.Conclusion:In general,the health status of the floating population in China is relatively good.The main influencing factors of health included gender and age.The central tendency of health inequity is reflected in the south-east coastal and northern regions,which are characterized by poverty.Attention to spatial aggregation is not only helpful to analyze the reasons of floating population,but also to study the health differences between different regions and health-related factors,to improve the overall health level of the whole population.

Objective To analyze the influencing factors of choroidopathy(choroidal atrophy and choroidal neovas-cularization)secondary to high myopia based on Logistic regression analysis and to construct a Nomogram risk prediction model based on the related factors,so as to provide guidance for clinical treatment.Methods A total of 340 patients(680 eyes)with high myopia admitted to Beijing Jishuitan Hospital from January 2021 to January 2023 were selected and di-vided into group A(170 patients,340 eyes)and group B(170 patients,340 eyes).The incidence of choroidopathy in the two groups was compared.The groups A and B were divided into two subgroups,subgroup a and subgroup b,according to whether choroidopathy occurred or not.Multivariate Logistic regression analysis was carried out to explore the influencing factors of choroidopathy secondary to high myopia.A Nomogram risk prediction model for choroidopathy secondary to high myopia was constructed based on the influencing factors and externally validated.Results In groups A and B,the age,proportion of diabetes mellitus,axial length,and level of seruim transforming growth factor β1(TGF-β1)of patients in subgroup a were higher than those in the subgroup b,and the diopter was lower than that in the subgroup b(all P＜0.05).The Logistic regression analysis showed that age,diabetes mellitus,axial length and serum TGF-β1 level were independent risk factors for choroidopathy secondary to high myopia,and diopter was a protective factor(all P＜0.05).Age,diabetes mellitus,axial length and serum TGF-β1 level were positively correlated risk factors for choroidopathy secondary to high myopia,and diopter was a negatively correlated risk factor(all P＜0.05).The area under the curve of the Nomogram risk prediction model for predicting choroidopathy secondary to high myopia was 0.818,and the calibration was good.Con-clusion Age,diabetes mellitus,axial length,diopter and serum TGF-β1 level are the influential factors for choroidopa-thy secondary to high myopia.The Nomogram risk prediction model established based on these factors has a certain value for predicting choroidopathy secondary to high myopia.The clinical therapeutic schedules should be made based on this model to reduce the risk of secondary choroidopathy.

Objective To analyze the latent profiles of active aging among community older adults and its influencing factors,and explore relationship of different categories and nursing care needs.Methods Multistage stratified random sampling was used to select 341 community older people in Qingdao from June 2023 to August 2023 as the survey population.The study instruments included the General Information Questionnaire,the Active Ageing Scale,the Care Needs of the Elderly Scale.Latent profile analysis was used to explore the latent profiles of active aging on the community older adults.The influencing factors of latent profiles were identified by multivariate Logistic regression.Results The active aging of the community older people was identified as a model with 3 latent categories,defined as low active aging type(38.l％),medium active aging-high spiritual intelligence type type(51.6％),and high active aging-comprehensive type(10.3％).The influencing factors include age,education,monthly income level,nursing needs(all P＜0.05).Community older adults with low self-development and contribution needs are more likely to be low active aging type.Community older adults with high needs for life care and living environment(OR=3.268,P=0.0l 1),physical and psychological support(OR=1.972,P=0.025),and interpersonal communication and health knowledge(OR=3.433,P＜0.001)were more likely to be medium active aging-high spiritual intelligence type.Those with low protection and security needs(OR=0.446,P=0.012)were more likely to be medium active aging-high spiritual intelligence type type.Community older adults with low health monitoring needs(OR=0.297,P=0.029)and high specialty care needs(OR=3.019,P=0.033)were more likely to be high active aging-comprehensive type.Conclusion The level of active aging of older adults in the community is medium,which is characterized by 3 categories.Community nursing staffs should focus on the elderly with low active aging type and medium active aging-high spiritual intelligence type,and targeted intervention should be adopted according to different category characteristics,so as to accurately meet their nursing needs,finally improve the level of active aging of community older adults.

In this study, a new in situ film of compound iodine oral spray was prepared by in situ gel technology, which was used to exert sustained-release effect for promoting the repair of oral ulcer wounds. Firstly, the formulation and process of the spray solution were optimized according to the spray state and film-forming time. The drug-liquid mixing ratio was evaluated by film-forming time and drug film adhesion. The drug content, stability, pH, and spraying effect of compound iodine oral spray prepared by the optimal formulation were investigated; and the physicochemical properties, including film formation time, solubility, hygroscopicity, moisture retention and in vitro release of drug film were evaluated. In addition, the biocompatibility of the film-forming materials and proliferation ability of drug film were investigated by cell experiment. Through the rabbit oral ulcer model, the in vivo film-forming and repair-promoting effects of compound iodine oral spray were evaluated. The results showed that the pH of liquid A and liquid B prepared were 6.21±0.02 and 6.42±0.03, respectively, which were in line with the normal pH range of oral mucosa; liquid A and liquid B had good stability and spray state; the iodine content in solution B was (1.96±0.01) mg/mL; the in situ membrane formation time in vitro and in the oral cavity were (118.3±3.6) s and (133.3±4.6) s, respectively; the 24-hour dissolution rate was (87.31±1.74)%, the moisture absorption rate was (124.17±7.13)%, and the moisture retention rate was (26.85±2.50)%; the iodine content in the oral spray was (47.42±0.39) mg/g, with good flexibility and adhesion, as well as some slow-release effect. In cell experiment, the film-forming materials showed good biocompatibility and growth promotion ability. The results of the rabbit oral ulcer experiment showed that the compound iodine oral spray could rapidly form a film in vivo and significantly promote the repair of oral ulcer. In conclusion, the compound iodine oral spray in situ film with a stable preparation process can effectively promote the repair of oral ulcer wounds, which provides a new idea for the research of novel oral mucosa formulations, with a good prospect of transfer.