Fungal infections have emerged as a critical public health issue endangering human health. However, the existing arsenal of antifungal agents is limited in diversity and is commonly plagued by drawbacks including narrow antimicrobial spectrums and the frequent emergence of drug resistance, which severely compromises the efficacy of clinical treatments. Pathogenic fungi can develop extensive adaptability to currently available drugs through multiple mechanisms, which are mainly manifested in three aspects: drug resistance, tolerance and persistence. The molecular mechanisms and regulatory pathways underlying drug resistance, tolerance and persistence in pathogenic fungi were systematically summarized in this review, and the counteractive strategies such as combination therapy and the development of novel antifungal agents were further discussed, which aimed to provide theoretical basis and practical reference for the precision treatment of fungal infections.

BACKGROUND:Our previous study found that Modified Erxian Pill could alleviate inflammation in collagen-induced arthritis rats,but its mechanism needs to be further verified. OBJECTIVE:To analyze the components absorbed in the blood of Modified Erxian Pill,and observe the effect of the drug-containing serum of Modified Erxian Pill on pyroptosis of J774A.1 macrophages. METHODS:(1)Analysis of components absorbed in the blood of Modified Erxian Pill:Ultra-high performance liquid chromatography-high resolution mass spectrometry was used to detect and identify Modified Erxian Pill and its components absorbed in the blood.(2)Effect of the drug-containing serum of Modified Erxian Pill on pyroptosis of J774A.1 macrophages:Molecular docking technology was used to initially verify the sesquiterpenoids and NLRP3 in components absorbed in the blood of Modified Erxian Pill.J774A.1 macrophages were randomly divided into blank control group,lipopolysaccharide+adenosine triphosphate group,and lipopolysaccharide+adenosine triphosphate+Modified Erxian Pill with low(2.5%),medium(5%),and high(10%)dose groups.The release of lactate dehydrogenase in the cell supernatant of each group was detected according to the kit instructions.The levels of interleukin-1β and interleukin-18 in cell supernatant were detected in each group by ELISA.The cell membrane damage was detected by Hoechst/PI staining.The expression levels of NLRP3,Caspase-1,GSDMD,and GSDMD-N protein in the cells of each group were detected by western blot assay. RESULTS AND CONCLUSION:(1)A total of 32 active components of Modified Erxian Pill were identified,and 21 components entered the blood.The main components into blood included a variety of sesquiterpenoids.(2)Molecular docking results showed that 3-O-Acetyl-13-deoxyphomenone,Incensol oxide,Atractylenolide III,Rupestonic acid,and 3,7-Dihydroxy-9,11-eremophiladien-8-one had good binding activity with NLRP3.(3)Compared with the blank control group,lactate dehydrogenase activity and the expression levels of interleukin-1β and interleukin-18 were significantly increased in cell supernatant of lipopolysaccharide+adenosine triphosphate group(P<0.001).Hoechst/PI staining showed that the number of PI-positive cells was significantly increased.After the intervention of lipopolysaccharide+adenosine triphosphate+Modified Erxian Pill group,all of them showed different degrees of reduction.(4)Compared with the blank control group,NLRP3,Caspase-1,GSDMD,and GSDMD-N protein expression levels were significantly increased in the lipopolysaccharide+adenosine triphosphate group(P<0.05).Compared with lipopolysaccharide+adenosine triphosphate group,the protein expressions of NLRP3,Caspase-1,GSDMD,and GSDMD-N were significantly decreased in the lipopolysaccharide+adenosine triphosphate+Modified Erxian Pill group(P<0.05),and had a certain dose dependence.These findings verify that the drug-containing serum of Modified Erxian Pill may inhibit the pyroptosis of J774A.1 macrophages by regulating the NLRP3/Caspase-1/GSDMD pathway.

Endoplasmic reticulum stress(ERS)plays a crucial role in cellular self-protection.In response to environmental changes,cells activate the unfolded protein response(UPR)and endoplasmic reticulum-associated protein degradation pathways to restore homeostasis.Increasing experimental evidence indicates that hypoxia and inflammation negatively impact synoviocytes in rheumatoid arthritis(RA),leading to the activation of three key endoplasmic reticulum transmembrane proteins:inositol-requiring enzyme 1,protein kinase R-like endoplasmic reticulum kinase,and activating transcription factor 6.These proteins mediate UPR pathways that are implicated in the pathological progression of RA.Targeting ERS-related molecules has emerged as a promising therapeutic strategy for RA.This article reviews the impact of ERS on the pathogenesis of RA and discusses drugs that modulate ERS as potential treatments for the disease.

Objective:To explore the prominent manifestations and internal associations of the overuse of short-form videos,inattention,and school refusal behavior in junior high school students.Methods:A survey was conducted among 1 106 junior high school students,utilizing the Short-form Video APP Overuse Scale,the Inatten-tion subscale of the Swanson,Nolan,and Pelham Rating Scale(SNAP-Ⅳ),and the School Refusal Behavior Ques-tionnaire to assess the extent of short-form video overuse,inattention,and school refusal behavior.By using the R language for network analysis,the core nodes of individual networks and the bridge nodes connecting the combined networks were analyzed,and the structural differences in networks across genders and grades were compared.Re-sults:The core nodes of the network pertaining to overuse of short-form video,inattention,and school refusal behav-ior were"Using short videos enhances connection with others"(expected influence value=1.03),"easily distract-ed"(expected influence value=1.99),and"school alienation"(expected influence value=0.83).The bridge nodes in the combined network were"defiant behavior"(bridge expected influence value=0.69)and"difficulty sustaining mental activities"(bridge expected influence value=0.47).Analysis of network structure differences i-dentified that the core nodes within the combined networks of the first,second,and third grades of junior high school students were respectively"prioritizing short videos over other activities","defiant behavior",and"academic aptitude".Conclusion:Junior high school students exhibiting overuse of short videos,inattention,or school refusal behavior are characterized by enhanced social connectivity through platforms such as short video apps,a propensity for distraction,and feelings of school alienation,respectively.When these three behaviors coexist,the most salient related factor is oppositional behavior,coupled with difficulties in sustaining cognitive tasks and maintaining con-centration.

Objective:To evaluate the effect of low-dose esketamine on intraoperative electroencephalogram (EEG) burst suppression under general anesthesia in elderly patients.Methods:In this prospective randomized controlled trial, 86 elderly patients of either sex, scheduled for elective lumbar fusion surgery at Anhui No. 2 Provincial People′s Hospital from March 15 to June 1, 2024, aged 60-85 yr, with a body mass index of 18-28 kg/m 2, of American Society of Anesthesiologists Physical Status classification ⅡorⅢ, were divided into 2 groups ( n=43 each) using a random number table method: esketamine group (group S) and control group (group C). In group S, esketamine at a dose of 0.25 mg/kg was intravenously injected after anesthesia induction, while the equal volume of normal saline was intravenously injected in group C. The occurrence of EEG burst suppression and frequency of the characteristic density spectral array were recorded using the Masimo Sedline cerebral function monitor. The patient state index was recorded before esketamine administration and at 5, 10, 30 and 60 min after administration and at 5 min before the end of the surgery. Delirium was assessed using the Confusion Assessment Method at 1 and 3 days after operation. The total pressing times of patient-controlled analgesia within 48 h after operation, requirement for rescue analgesia and occurrence of adverse reactions (nausea and vomiting, delayed emergence from anesthesia, agitation, headache) were recorded. The consumption of ephedrine, phenylephrine, sufentanil and remifentanil during operation, emergence time, time of tracheal extubation, length of stay in the post-anesthesia care unit, and length of postoperative hospital stay were also recorded. Results:Compared with group C, the incidence of EEG burst suppression and frequency of the characteristic density spectral array were significantly decreased, the patient state index values were increased at each time point after administration and 5 min before the end of surgery, the consumption of ephedrine, phenylephrine, sufentanil and remifentanil during operation was reduced, the effective pressing times of patient-controlled analgesia, rate of rescue analgesia, and incidence of postoperative delirium within 3 days after operation were decreased, and the length of postoperative hospital stay was shortened in group S ( P<0.05). Conclusions:Low-dose esketamine can reduce intraoperative EEG burst suppression and decrease the development of postoperative delirium under general anesthesia in elderly patients.

BACKGROUND:Polyamines play a crucial role in tissue calcification.Spermidine/spermine N1-acetyltransferase 1(SAT1),as a key rate-limiting enzyme regulating intracellular polyamine metabolism,has been associated with various pathological processes.However,its role in vascular calcification remains unclear.OBJECTIVE:To investigate the role of SAT1 in rat vascular smooth muscle cell calcification.METHODS:(1)Bioinformatics analysis:Differential expression of SAT1 in human carotid atherosclerotic plaques and their surrounding healthy carotid artery tissues were using GEO datasets.PanglaoDB database was used to analyze SAT1 expression abundance and localization across different cell types through single-cell sequencing.(2)Rat vascular smooth muscle cells were divided into three groups:a control group cultured in DMEM medium,a calcification group induced by DMEM medium containing 10 mmol/L β-glycerophosphate sodium and 3 mmol/L calcium chloride,and the 50,100 μmol/L diacetylaminotriazamidine groups treated with the SAT1 inhibitor,diacetylaminotriazamidine,in addition to the calcification medium.After 7-10 days of culture,alizarin red S staining was performed,and cellular calcium content and alkaline phosphatase activity were assessed.Western blot was used to detect the protein expression of Runt-related transcription factor 2,bone morphogenetic protein 2,alpha-smooth muscle actin,and SAT1.Immunofluorescence staining was conducted to examine the expression of Runt-related transcription factor 2 and SAT1.RESULTS AND CONCLUSION:(1)Bioinformatics analysis revealed significantly upregulated expression of SAT1 and Runt-related transcription factor 2(P<0.05)in carotid atherosclerotic plaques compared with healthy carotid tissues(P<0.05).Single-cell sequencing database analysis confirmed SAT1 expression in vascular smooth muscle cells.(2)Compared with the control group,the calcification group showed significantly increased Runt-related transcription factor 2,bone morphogenetic protein 2,SAT1,calcium content,and alkaline phosphatase activity,while alpha-smooth muscle actin expression was significantly decreased(all P<0.05).Compared with the calcification group,the 50 and 100 μmol/L diacetylaminotriazamidine groups showed significantly decreased Runt-related transcription factor 2,bone morphogenetic protein 2,calcium content,and alkaline phosphatase activity,while alpha-smooth muscle actin expression was significantly increased(all P<0.05).(3)Immunofluorescence experiments demonstrated that compared with the calcification group,the expression intensity of Runt-related transcription factor 2 was significantly reduced in the 50 and 100 μmol/L diacetylaminotriazamidine groups.Overall,SAT1 may promote vascular smooth muscle cell calcification by upregulating Runt-related transcription factor 2 expression.

Objective:To explore the prominent manifestations and internal associations of the overuse of short-form videos,inattention,and school refusal behavior in junior high school students.Methods:A survey was conducted among 1 106 junior high school students,utilizing the Short-form Video APP Overuse Scale,the Inatten-tion subscale of the Swanson,Nolan,and Pelham Rating Scale(SNAP-Ⅳ),and the School Refusal Behavior Ques-tionnaire to assess the extent of short-form video overuse,inattention,and school refusal behavior.By using the R language for network analysis,the core nodes of individual networks and the bridge nodes connecting the combined networks were analyzed,and the structural differences in networks across genders and grades were compared.Re-sults:The core nodes of the network pertaining to overuse of short-form video,inattention,and school refusal behav-ior were"Using short videos enhances connection with others"(expected influence value=1.03),"easily distract-ed"(expected influence value=1.99),and"school alienation"(expected influence value=0.83).The bridge nodes in the combined network were"defiant behavior"(bridge expected influence value=0.69)and"difficulty sustaining mental activities"(bridge expected influence value=0.47).Analysis of network structure differences i-dentified that the core nodes within the combined networks of the first,second,and third grades of junior high school students were respectively"prioritizing short videos over other activities","defiant behavior",and"academic aptitude".Conclusion:Junior high school students exhibiting overuse of short videos,inattention,or school refusal behavior are characterized by enhanced social connectivity through platforms such as short video apps,a propensity for distraction,and feelings of school alienation,respectively.When these three behaviors coexist,the most salient related factor is oppositional behavior,coupled with difficulties in sustaining cognitive tasks and maintaining con-centration.

Objective:To investigate the level of rehabilitation exercise adherence in patients undergoing anterior cruciate ligament (ACL) reconstruction within 6 months post-surgery, analyze its trajectory, and provide a reference for clinical nursing interventions.Methods:Using a convenience sampling method, 150 patients who underwent arthroscopic ACL reconstruction at the First Affiliated Hospital of Zhengzhou University from June 2022 to August 2023 were selected. Rehabilitation exercise adherence was assessed monthly from postoperative 1 month (M 1) to 6 months (M 6) using the Orthopedic Functional Exercise Adherence Scale. A latent class growth model was employed to analyze adherence trajectory categories, and Logistic regression was used to identify influencing factors for different adherence trajectories. Results:A total of 144 patients completed follow-up (follow-up rate was 96.00%). Exercise adherence scores gradually declined over the 6 months post-surgery, with scores of (50.30±5.39), (45.92±3.85), (39.48±5.27), (35.83±6.19), (31.85±7.59), and (29.88±7.88) from M 1 to M 6, respectively. The difference in scores across time points was statistically significant ( F=85.467, P<0.01). Three potential categories were identified through the latent class growth model: the moderate-high level rapid decline group ( n=65, 45.14%), the moderate-low level stable group ( n=32, 22.22%), and the moderate-low level slow decline group ( n=47, 32.64%). Logistic regression analysis revealed that gender and age were significant factors influencing adherence trajectory categories ( P<0.05) . Conclusions:Dynamic declines in rehabilitation exercise adherence are common among ACL reconstruction patients, with varying degrees of decline. Gender and age may influence adherence trajectories, suggesting that targeted management strategies could be implemented in clinical practice.

BACKGROUND:Rheumatoid arthritis is an inflammatory disease.Many studies have shown that wogonin has a good anti-inflammatory effect on rheumatoid arthritis,but its exact efficacy and specific mechanism of action remain to be clarified. OBJECTIVE:To investigate the mechanism of wogonin ameliorating joint inflammation by regulating endoplasmic reticulum stress pathway in rats with collagen-induced arthritis. METHODS:(1)At the animal level:Female Wistar rats were divided into healthy control group,arthritis model group and wogonin treatment group.Rat models of arthritis in the latter two groups were established by subcutaneous injection of bovine type Ⅱ collagen and adjuvant.In the wogonin group,wogonin was given by gavage for 28 consecutive days after modeling.During this period,the rats in each group were weighed,and arthritis score and ankle swelling were measured every 7 days.After the experiment,the pathological changes of the joint were observed,the mRNA and protein levels of endoplasmic reticulum stress pathway GRP78 and CHOP were detected by qRT-PCR,western blot,and immunohistochemistry.(2)At the cellular level,cell counting kit-8 was used to detect the cytotoxic effect of wogonin on fibroblast-like synoviocytes from rats with collagen-induced arthritis.The fibroblast-like synoviocytes induced by thapsigargin were treated with different concentrations of wogonin.The levels of interleukin-1β and tumor necrosis factor-α in the cell supernatant were detected by ELISA,and the intracellular reactive oxygen species in each group were determined by DCFH-DA probe method.The mRNA and protein levels of GRP78,IRE1α,XBP1s and CHOP were detected by qRT-PCR and western blot,respectively. RESULTS AND CONCLUSION:Compared with the healthy control group,arthritis index score and ankle swelling degree in the arthritis model group were increased(P<0.01),synovial hyperplasia,inflammatory cell infiltration,cartilage destruction and bone erosion were observed in pathological sections,and the mRNA and protein expressions of GRP78 and CHOP in the ankle were significantly increased(P<0.01),which were mainly located in synovial tissue and articular surface.Compared with the arthritis model group,the arthritis index score and ankle swelling degree in the wogonin treatment group were decreased(P<0.05),synovial hyperplasia and the number of inflammatory cells were decreased,cartilage destruction and bone erosion were alleviated,the mRNA and protein expression levels of GRP78 and CHOP in the ankle were decreased(P<0.05),particularly in synovial tissue and on the articular surface.There was no significant difference in body mass among the three groups(P>0.05).In the cell experiment,200 μmol/L wogonin significantly reduced the survival rate of fibroblast-like synoviocytes(P<0.01).Compared with the blank control group,the levels of interleukin-1β,tumor necrosis factor-α,content of reactive oxygen species,and mRNA and protein expression of GRP78,IRE1α,XBP1s,and CHOP in the thapsigargin group were significantly increased(P<0.05);compared with the thapsigargin group,50 and 100 μmol/L wogonin significantly reduced the levels of interleukin-1β and tumor necrosis factor-α in the cell supernatant(P<0.05,P<0.01),and 100 μmol/L wogonin significantly reduced the content of reactive oxygen species(P<0.01)and down-regulated the mRNA and protein expression levels of GRP78,IRE1α,XBP1s and CHOP(all P<0.05).These results suggest that wogonin can effectively alleviate joint inflammatory responses in rats with collagen-induced arthritis,and the endoplasmic reticulum stress pathway may be the key target of its intervention.

ObjectiveTo investigate the effects of Congrong Zonggan capsules （CRZG） on cognitive impairment in the Alzheimer's disease （AD） model of mice and its related mechanisms. MethodsSPF grade 4-week-old 5×FAD mice were divided into a model group， low-dose CRZG （0.819 g·kg^-1） and high-dose CRZG （1.638 g·kg^-1） groups， and Donepezilepezil hydrochloride group （2 mg·kg^-1）， with eight mice in each group. Eight C57 mice with the same background were set as the normal group. After one week of adaptive feeding， mice were orally administered continuously for six months. On the 5th month of drug administration， Y maze， new object recognition， and Morris water maze tests were conducted separately. After administration， mouse brain tissue was taken， and the levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in brain tissue were detected by enzyme-linked immunosorbent assay （ELISA）. Immunofluorescence （IF） was used to detect the expression of small glial cell markers Iba1， astrocyte markers GFAP， and amyloid protein 1-42 （Aβ_1-42） in the hippocampus of the brain tissue. The hematoxylin-eosin （HE） staining was used to detect pathological changes in the hippocampus of brain tissue. Western blot was used to detect the expression of nuclear factor-κB （NF-κB） p65， NOD-like receptor protein 3 （NLRP3）， cleaved Caspase-1， apoptosis-associated speck-like protein containing a CARD （ASC）， and other proteins in the brain tissue. ResultsCompared with those in the normal group， the mice in the model group had obvious cognitive impairment. The spontaneous alternation rate of the Y maze was decreased， and the discrimination index of novel object recognition was decreased significantly （P<0.01）. The escape latency in the water maze was shortened significantly （P<0.01）. The contents of IL-6 and TNF-α in brain tissue were increased. The fluorescence levels of Iba1 and Aβ_1-42 in the hippocampus were significantly increased （P<0.01）. There was a significant increase in neuronal lesions， neuronal atrophy， loose arrangement of tissue structure， and abnormal erythrocyte aggregation in the hippocampus. The protein expressions of p-NF-κB p65/NF-κB p65， cleaved Caspase-1， ASC， IL-6， and IL-1β were significantly increased （P<0.05， P<0.01）. Compared with the model group， the spontaneous alternation rate and discrimination index of the high-dose CRZG group were increased significantly （P<0.01）， and the escape latency was shortened significantly （P<0.05， P<0.01）. The content of IL-6 decreased in the brain， and that of TNF-α dropped significantly （P<0.01）. The expression of Iba1 protein and Aβ_1-42 in the hippocampus decreased significantly （P<0.05， P<0.01）. The hippocampal neurons were densely arranged， and the pyramidal nuclei were clear and centered. The abnormal aggregation of red blood cells was alleviated. The value of p-NF-κB/NF-κB proteins and the expression of ASC， cleaved Caspase-1， IL-6， and IL-1β were significantly decreased （P<0.05， P<0.01）. ConclusionCRZG can effectively improve cognitive impairment in 5×FAD mice with Alzheimer's disease， and its mechanism may be related to the regulation of the NF-κB/NLRP3 pathway to reduce the abnormal activation of microglia and inhibit neuroinflammation.