BACKGROUND:Due to the aging population and the increase in elderly diseases,research and utilization of epidermal stem cells in the elderly have become increasingly important.However,due to the weak proliferation and drying ability of epidermal stem cells in the elderly,their application in biomedical and clinical research is limited.OBJECTIVE:To investigate the ability of rotating cell culture system to promote the proliferation and maintenance of epidermal stem cells in elderly individuals.METHODS:Epidermal stem cells were obtained from children (7-12 years old) and the elderly (over 60 years old) using traditional planar culture methods.After identification by cell morphology and cell immunofluorescence,the differences in proliferation ability of epidermal stem cells between the children and the elderly were analyzed by flow cytometry,CCK-8 assay,and cell clone formation assay.Elderly epidermal stem cells were cultured using a rotating cell culture system and 3D TableTrix? microcarriers.The cell proliferation and expression of stemness markers were detected by Live/Dead staining,cell doubling efficiency,cell doubling time,and real-time quantitative polymerase chain reaction.RESULTS AND CONCLUSION:(1) Under planar culture conditions,the children group had stronger proliferative ability than the elderly group.(2) Compared with planar culture,the dynamic cultivation of elderly epidermal stem cells using a rotating cell culture system had a higher population doubling (P<0.01) and a shorter population doubling time (P<0.01).(3) The dynamic cultivation of rotating cell culture system promoted the self-assembly of elderly epidermal stem cells to form 3D human epidermal stem cell spheres based on microcarrier scaffolds,which had similar epidermal stem cell proliferation as the children group.(4) After dynamic culture,the expression of stemness marker genes (ITGA6 and ITGB1),basal cell marker genes (K5) and differentiation marker genes (K10 and K1) of elderly epidermal stem cells reached the same level as that of children.The results showed that the dynamic cultivation of rotating cell culture system can not only improve the efficiency of elderly epidermal stem cell culture,promote cell proliferation and self-assembly into 3D cell spheres,but also maintain a certain stemness of elderly epidermal stem cells without complete terminal differentiation.

AIM:To investigate the ameliorative effect of interleukin 17A(IL-17A)gene knockout on airway remodeling in asthmatic mice and the specific mechanism involved.METHODS:In vivo,wild-type and IL-17A knockout(IL-17A-/-)mice were divided into 4 groups:control,IL-17A-/-,ovalbumin(OVA)and OVA+IL-17A-/-,with 6 mice in each group.Airway hyperresponsiveness was measured,pathological changes in the lungs were observed by staining tissue sections,the expression of the proteins related to airway remodeling,such as α-smooth muscle actin(α-SMA),matrix me-talloproteinase-9(MMP-9)and collagen type Ⅲ(Col Ⅲ),was detected by immunohistochemistry,and the level of p-p38 mitogen-activated protein kinase was detected by Western blot.In vitro,the regulatory effects of IL-17A on transforming growth factor β1(TGF-β1)-induced proliferation and migration of human airway smooth muscle(HASM)cells were as-sessed.Additionally,the level of p38 phosphorylation was detected,and the effect of a p38 inhibitor on airway remodel-ing-related protein expression was evaluated.RESULTS:Knockout of IL-17A significantly attenuated airway hyperrespon-siveness,airway inflammation and downregulated α-SMA(P<0.01),MMP-9(P<0.01)and Col Ⅲ(P<0.01)in asth-matic mice.In HASM cells,the proliferation and migration of these cells were inhibited following IL-17A intervention.Furthermore,a significant reduction in p38 phosphorylation was observed in both mouse lung tissue(P<0.01)and HASM cells(P<0.01).In addition,the gene expression of α-SMA(P<0.05),MMP-9(P<0.01)and Col Ⅲ(P<0.05)was re-duced upon further inhibition of p38 phosphorylation.CONCLUSION:Knockout of IL-17A attenuates airway remodeling in asthmatic mice by inhibiting the phosphorylation of p38 in airway smooth muscle.

An increasing number of studies have shown that there is a close relationship between Alzheimer's disease(AD)and diabetes mellitus(DM).Traditional Chinese medicine holds that"kidney yin deficiency"is the common pathogenesis of these two diseases,while modern medicine believes that their pathogenesis involves abnormal aggregation of amyloid proteins,insulin deficiency and resistance,inflammatory response,oxidative stress,and autophagy,among others.The well-known traditional Chinese medicine formula Liuwei Dihuang Pills plays a significant role in the treatment of these two diseases with the same therapeutic approach.Therefore,this article will explore the connection between Liuwei Dihuang Pills and the treatment of AD and DM from different aspects;analyze the common etiology and pathogenesis of AD and DM,and explain the mechanism of prevention and treatment of Liuwei Dihuang Pills,with the aim of providing new ideas and methods for the integrated traditional and Western medicine prevention and treatment of AD and DM in the future.

BACKGROUND:Due to the aging population and the increase in elderly diseases,research and utilization of epidermal stem cells in the elderly have become increasingly important.However,due to the weak proliferation and drying ability of epidermal stem cells in the elderly,their application in biomedical and clinical research is limited.OBJECTIVE:To investigate the ability of rotating cell culture system to promote the proliferation and maintenance of epidermal stem cells in elderly individuals.METHODS:Epidermal stem cells were obtained from children (7-12 years old) and the elderly (over 60 years old) using traditional planar culture methods.After identification by cell morphology and cell immunofluorescence,the differences in proliferation ability of epidermal stem cells between the children and the elderly were analyzed by flow cytometry,CCK-8 assay,and cell clone formation assay.Elderly epidermal stem cells were cultured using a rotating cell culture system and 3D TableTrix? microcarriers.The cell proliferation and expression of stemness markers were detected by Live/Dead staining,cell doubling efficiency,cell doubling time,and real-time quantitative polymerase chain reaction.RESULTS AND CONCLUSION:(1) Under planar culture conditions,the children group had stronger proliferative ability than the elderly group.(2) Compared with planar culture,the dynamic cultivation of elderly epidermal stem cells using a rotating cell culture system had a higher population doubling (P<0.01) and a shorter population doubling time (P<0.01).(3) The dynamic cultivation of rotating cell culture system promoted the self-assembly of elderly epidermal stem cells to form 3D human epidermal stem cell spheres based on microcarrier scaffolds,which had similar epidermal stem cell proliferation as the children group.(4) After dynamic culture,the expression of stemness marker genes (ITGA6 and ITGB1),basal cell marker genes (K5) and differentiation marker genes (K10 and K1) of elderly epidermal stem cells reached the same level as that of children.The results showed that the dynamic cultivation of rotating cell culture system can not only improve the efficiency of elderly epidermal stem cell culture,promote cell proliferation and self-assembly into 3D cell spheres,but also maintain a certain stemness of elderly epidermal stem cells without complete terminal differentiation.

An increasing number of studies have shown that there is a close relationship between Alzheimer's disease(AD)and diabetes mellitus(DM).Traditional Chinese medicine holds that"kidney yin deficiency"is the common pathogenesis of these two diseases,while modern medicine believes that their pathogenesis involves abnormal aggregation of amyloid proteins,insulin deficiency and resistance,inflammatory response,oxidative stress,and autophagy,among others.The well-known traditional Chinese medicine formula Liuwei Dihuang Pills plays a significant role in the treatment of these two diseases with the same therapeutic approach.Therefore,this article will explore the connection between Liuwei Dihuang Pills and the treatment of AD and DM from different aspects;analyze the common etiology and pathogenesis of AD and DM,and explain the mechanism of prevention and treatment of Liuwei Dihuang Pills,with the aim of providing new ideas and methods for the integrated traditional and Western medicine prevention and treatment of AD and DM in the future.

AIM:To investigate the ameliorative effect of interleukin 17A(IL-17A)gene knockout on airway remodeling in asthmatic mice and the specific mechanism involved.METHODS:In vivo,wild-type and IL-17A knockout(IL-17A-/-)mice were divided into 4 groups:control,IL-17A-/-,ovalbumin(OVA)and OVA+IL-17A-/-,with 6 mice in each group.Airway hyperresponsiveness was measured,pathological changes in the lungs were observed by staining tissue sections,the expression of the proteins related to airway remodeling,such as α-smooth muscle actin(α-SMA),matrix me-talloproteinase-9(MMP-9)and collagen type Ⅲ(Col Ⅲ),was detected by immunohistochemistry,and the level of p-p38 mitogen-activated protein kinase was detected by Western blot.In vitro,the regulatory effects of IL-17A on transforming growth factor β1(TGF-β1)-induced proliferation and migration of human airway smooth muscle(HASM)cells were as-sessed.Additionally,the level of p38 phosphorylation was detected,and the effect of a p38 inhibitor on airway remodel-ing-related protein expression was evaluated.RESULTS:Knockout of IL-17A significantly attenuated airway hyperrespon-siveness,airway inflammation and downregulated α-SMA(P<0.01),MMP-9(P<0.01)and Col Ⅲ(P<0.01)in asth-matic mice.In HASM cells,the proliferation and migration of these cells were inhibited following IL-17A intervention.Furthermore,a significant reduction in p38 phosphorylation was observed in both mouse lung tissue(P<0.01)and HASM cells(P<0.01).In addition,the gene expression of α-SMA(P<0.05),MMP-9(P<0.01)and Col Ⅲ(P<0.05)was re-duced upon further inhibition of p38 phosphorylation.CONCLUSION:Knockout of IL-17A attenuates airway remodeling in asthmatic mice by inhibiting the phosphorylation of p38 in airway smooth muscle.

Objective: To understand the prevalence of myopia among primary and secondary school students in minority areas of Yunnan Province, and to explore the influence of outdoor activities and short-distance use of eyes, so as to provide a basis for early myopia intervention. Methods: In October 2020, the survey was conducted among 1 782 primary and secondary school students in three cities of Yunnan through a multi stage random cluster sampling method. All subjects underwent a questionnaire survey and the visual acuity examination at baseline. The first follow-up was conducted in October 2021 to obtain 1 691 valid samples, and the second follow-up was conducted in May 2023 to obtain 1 367 valid samples. Factors associated with myopia among primary and secondary school students were explored by using generalized estimating equations. Results: The prevalence rates of myopia in 2020, 2021 and 2023 were 52.64%, 61.62% and 69.35%, respectively, showing an increasing trend ( χ 2 trend =91.77, P <0.05). The results of multivariate regression analysis on the generalized estimation equations showed that age at baseline ( OR =1.31), girls ( OR =1.76), Hani ethnicity ( OR =0.75), Bai ethnicity ( OR =0.69), parental myopia ( OR =1.97-2.29), parents often reducing children s exercise time for homework or tutoring ( OR =1.35), less than 1 time or 2-3 times of ball sports per week ( OR =1.27, 1.20 ), reading and writing in the classroom during the break ( OR =1.27), reading in direct sunlight occasionally ( OR =1.20), using only desk lamp for writing at home ( OR =0.71), more than 1 hours of short-distance eye use for a break once ( OR =1.23) were associated with myopia among primary and secondary school students ( P <0.05). Conclusions Outdoor activities and short-distance use of eye among primary and middle school students in minority areas in Yunnan province are suboptimal. Enhancing the related environmental and behavioral factors can effectively mitigate the occurrence and progression of myopia.

Objective:This study aims to investigate the differences in emotional status and cognitive function among patients with vestibular migraine (VM), Meniere′s disease (MD), and their comorbidity (VMMD), and to analyze key factors influencing cognitive function.Methods:This cross-sectional study included 96 outpatients (32 males, 64 females, aged 21-73 years) from the Department of Otorhinolaryngology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, between December 2022 and December 2023. The study population consisted of 31 VM patients (VM group), 36 MD patients (MD group), and 29 VMMD patients (VMMD group), along with 32 healthy controls (16 males, 16 females, aged 19-74 years). Cognitive function was assessed using the Montreal Cognitive Assessment (MoCA), while emotional status and somatization symptoms were evaluated through the Generalized Anxiety Disorder scale, Patient Health Questionnaire Depression scale, Symptom Checklist-90, and the Self-rating Somatization Symptom scale. Multiple linear regression analysis was conducted to explore the influence of different variables on cognitive function.Results:The total MoCA score in the VMMD group (26.0 [24.5, 28.0]) was significantly lower than that in the control group (28.0 [27.0, 29.0]) and the MD group (28.0 [26.0, 30.0]) ( P=0.006). VMMD patients exhibited significant impairments in specific cognitive domains, including visuospatial/executive function, delayed recall, and orientation ( P<0.05). Patients with VM, MD, and VMMD showed higher rates of anxiety, depression, and somatization symptoms compared to the control group ( P<0.05), with the VMMD group experiencing the most severe emotional distress. Multiple linear regression analysis identified education level and vestibular disease type as key factors affecting cognitive function, with a university-level education predicting higher MoCA scores ( P<0.001), while VMMD was associated with cognitive decline ( P<0.01). Conclusions:Patients with VM and MD, particularly those with comorbid VMMD, exhibit significant emotional distress. Cognitive impairments are present in VM and VMMD patients, affecting different cognitive domains. These factors should be comprehensively considered in clinical assessments to develop more effective treatment strategies.

BACKGROUND:Currently,artificial corneas used for full-thickness transplantation lack biological activity and mechanical adaptability.Composite artificial corneas face interface issues between the corneal button and surrounding components.OBJECTIVE:To prepare an integrated artificial cornea with peptide enhancement,matched mechanical strength to natural cornea,and excellent transparency via in situ ultraviolet light curing of decellularized porcine cornea.METHODS:Non-ionic decellularization reagent Triton X-100 combined with ultrasonic freeze-thawing and super nucleases was utilized to prepare decellularized porcine cornea.Hydroxyethyl methacrylate monomer and photoinitiator were introduced into the decellularized porcine cornea simultaneously.Ultraviolet light with a filter was used to cover the peripheral region except for the central area,where polymerization was initiated using 275 nm ultraviolet light.After removal of unreacted monomers and initiators,the central optical zone was obtained.Similarly,the posterior lamellar layer was polymerized to form the hydrophobic barrier zone.Finally,REG active polypeptide was introduced to obtain in situ integrated full-layer artificial cornea.The physical properties,mechanical properties,transparency,degradation properties and in vivo and in vitro biocompatibility of artificial cornea were characterized.RESULTS AND CONCLUSION:(1)An optical region with the co-existence of polymer and collagen fibers was constructed in situ using hydroxyethyl methacrylate in the central region of decellularized porcine cornea.Under scanning electron microscopy,the upper surface of the artificial cornea was rough and irregular,with obvious concave and convex structure,and the lower surface was relatively smooth.The artificial cornea had mechanical properties close to those of natural cornea.The transparency of the optical zone reached 80%of that of the natural cornea.After soaking in PBS aseptic solution containing collagenase,it could preserve the solidified optical region and hydrophobic barrier zone,and maintain the basic structure of cornea.The artificial cornea had good cytocompatibility,could provide a suitable adhesion and growth environment for cells,was conducive to the migration and adhesion of corneal epithelial cells,promoted the growth of vascular endothelial cells and the formation of new blood vessels,and promoted the epithelialization process.The artificial cornea had good biocompatibility and safety after 12 weeks of subcutaneous implantation in SD rats,and could reduce the acute inflammatory reaction at the initial stage of implantation.(2)The results show that the integrated full-layer artificial cornea prepared by the experiment has the potential as a full-layer artificial cornea scaffold.

Objective:To investigate the impact of limonin(LIM)on the necrotic apoptosis of myocardial cells in rats with myocardial infarction(MI)by regulating the receptor-interacting protein 1(RIP1)/receptor-interacting protein 3(RIP3)/mixed-lineage kinase domain-like protein(MLKL)signaling pathway.Methods:A total of 60 Sprague-Dawley rats were randomly divided into sham-operation group(Sham group),MI model group(Model group),low-dose LIM group(LIM-L group,25 mg/kg LIM),high-dose LIM group(LIM-H group,50 mg/kg LIM),and high-dose LIM+RIP1 inhibitor Nec-1 group(LIM-H+Nec-1 group,50 mg/kg LIM+0.6 mg/kg Nec-1),with 12 rats in each group.A rat model of MI was established by ligation of the coronary artery.The changes of cardiac func-tion were examined for each group;HE staining was used to observe the pathological changes of myocardial tissue;the 2,3,5-triphen-yltetrazolium chloride-Evans blue method was used to measure myocardial infarct area;the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling method was used to observe the necrotic apoptosis of myocardial cells;quantitative real-time PCR and Western blot were used to measure the expression levels of mRNAs and proteins associated with the RIP1/RIP3/MLKL signaling pathway and the expression of apoptosis-related proteins.Results:Compared with the Sham group,the Model group had significant re-ductions in left ventricular ejection fraction,left ventricular systolic pressure,and the expression level of B-cell lymphoma-2(Bcl-2)in myocardial tissue(all P<0.001)and significant increases in left ventricular end-systolic volume,left ventricular end-diastolic pres-sure,left ventricular end-diastolic volume,myocardial infarct area,cell apoptosis rate,the mRNA and protein expression levels of RIP1,RIP3,and MLKL in myocardial tissue,and the protein expression levels of Bcl-2 associated X protein and cysteinyl aspartate-specific proteinase 3 in myocardial tissue(all P<0.001),as well as swelling and disordered arrangement of myocardial cells with ne-crosis and massive inflammatory cell infiltration on HE pathological sections.Compared with the Model group,the LIM-H group showed reverse changes in the above indicators(RIP1 mRNA:P=0.002,RIP3 mRNA:P=0.008,and the other indexes P were all<0.001),with alleviations of myocardial histopathological injury and inflammatory cell infiltration.Nec-1 promoted the effect of LIM in alleviating the necrotic apoptosis of myocardial cells in MI rats.Conclusion:LIM may alleviate the necrotic apoptosis of myocardial cells in MI rats by downregulating the RIP1/RIP3/MLKL signaling pathway.