Objective To investigate the role of poly(rC)-binding protein 1(PCBP1)in cadmium(Cd)-induced ferroptosis in mouse neuroblastoma Neuro-2a(N2A)cells.Methods N2A cells were exposed to a concentration gradient of CdCl?(0,1,2,4 μmol/L)for 72 h.Cell viability was assessed by trypan blue staining.Western blotting was employed to detect the expression of ferroptosis-related proteins(GPX4,HMOX1,ACSL4)and PCBP1.Intracellular Fe2? level and lipid peroxidation were detected using FerroOrange and BODIPY581/591 C11 probes,respectively.Ferrostatin-1(Fer-1),a ferroptosis inhibitor,was applied to confirm the critical role of ferroptosis in Cd-induced cytotoxicity.Molecular docking was performed to elucidate the interaction between PCBP1 and ferritin,as well as the binding sites of Cd2?.PCBP1 overexpression plasmid was further constructed for functional validation.Results Cd exposure suppressed cell viability in N2A cells in a dose-dependent manner(P<0.01),significantly down-regulated GPX4 expression(P<0.05),up-regulated HMOX1 expression(P<0.01),and induced Fe2? overload and lipid peroxidation(P<0.01).Molecular docking revealed that Cd2? directly bound to the KH2 domain of PCBP1 and then co-localized on the outer surface of ferritin heavy chain.Overexpression of PCBP1 markedly reversed Cd-induced Fe2? accumulation,GPX4 down-regulation,lipid peroxidation,and cell death.Conclusion Cd exposure disrupts PCBP1-mediated iron homeostasis via transcriptional suppression and competitive displacement of metal ions,and then synergistically drives Fe2? overload-triggered ferroptosis cascades,ultimately leading to neurotoxicity.Targeting PCBP1-mediated iron homeostasis can effectively mitigate Cd-induced neurotoxicity,and may serve as a novel therapeutic strategy.

Objective To explore serum levels of serine/threonine protein kinase 4(MST4)and heat shock protein 70(HSP70)in children with idiopathic immune thrombocytopenia(ITP)and their clinical signifi-cance.Methods Totally 98 children with ITP admitted to Northwest Women and Children's Hospital from April 2019 to April 2023 were retrospectively selected as the ITP group,and 50 healthy children who under-went physical examination during the same period were selected as the control group.Enzyme linked immu-nosorbent assay was used to detect serum levels of MST4 and HSP70,and the serum MST4 and HSP70 levels in children with different ITP levels were compared.The correlation between the indicators were analyzed by Pearson correlation.Logistic regression model was used to screen the prognostic factors of ITP,and the as-sessment value of serum MST4 and HSP70 on ITP prognosis was analyzed by subject working characteristic curve.Results Serum MST4,HSP70,and CD8 in the ITP group were higher than those in the control group,while PLT,CD3+,CD4+,CD4+/CD8+were lower than those in the control group,with statistical significance(P＜0.05).Serum MST4 and HSP70 levels in mild group,moderate group and severe group were increased successively,with statistical significance(P＜0.05).Correlation analysis showed that serum MST4 and HSP70 were positively correlated with CD8+(P＜0.05),and negatively correlated with PLT,CD3+,CD4+,CD4+/CD8+(P＜0.05).The disease course,serum MST4 and HSP70 of ITP children in the poor prognosis group were higher than those in the good prognosis group,and the differences were statistically significant(P＜0.05).Logistic regression analysis showed that the course of disease(OR=1.579,P＜0.001),serum MST4(OR=1.451,P＜0.001)and serum HSP70(OR=1.442,P＜0.001)were independent risk factors af-fecting the prognosis of children with ITP.The area under the curve of serum MST4 and HSP70 combined in the assessment of poor prognosis of ITP children was larger than that of serum MST4 and HSP70,and the difference was statistically significant(Z=4.568,4.672,both P＜0.001).Conclusion The elevated serum MST4 and HSP70 levels in children with ITP are related to the severity of the disease and cellular immune function.The combination of the two has a high evaluation value for the prognosis of children with ITP.

ObjectiveTo investigate the anti-inflammatory, antioxidant, and goblet cell-stimulating effects of a suspension of Ophiopogon japonicus (L. f.) Ker Gawl. (O. japonicus, Mai Dong) extract combined with hyaluronic acid (HA) in the mouse model with dry eye disease (DED).MethodsA DED mouse model was induced using benzalkonium chloride (BAK), followed by treatment with O. japonicus extract-containing eye drops at varying concentrations. Experimental groups included a normal control, a DED model control, a positive control, and an O. japonicus extract-treated group. Corneal fluorescein staining and tear break-up time (TBUT) were used to assess tear film stability and ocular surface integrity. Enzyme-linked immunosorbent assay (ELISA) measured inflammatory factor levels in corneal and conjunctival tissues, whereas Western blot (WB) analyzed key antioxidant and inflammatory markers, including nuclear factor erythroid 2-related factor (2Nrf2) and heme oxygenase 1 (HO-1). Periodic acid-schiff (PAS) staining and immunofluorescence were used to evaluate goblet cell density and mucin secretion.ResultsO. japonicus extract significantly improved corneal damage, reduced fluorescein staining scores, prolonged TBUT, and increased tear secretion. It downregulated inflammatory markers, including interleukin-8 (IL-8), interleukin-1β (IL-1β), and interferon-γ (IFN-γ) while upregulating Nrf2, HO-1, and the interleukin-13 (IL-13)/IFN-γ ratio, alleviating oxidative stress and inflammation. PAS staining showed increased conjunctival goblet cell density and restored mucin secretion, enhancing tear film stability.ConclusionO. japonicus extract demonstrated significant anti-inflammatory, antioxidant, and goblet cell-stimulating effects in a DED model, with good biocompatibility and promising therapeutic potential. Future research should optimize extraction processes and validate their efficacy and safety in clinical settings.

Medical institutions, with their clinical practice foundation and abundant human use experience data, have become important carriers for the inheritance and innovation of traditional Chinese medicine(TCM) and the "cradles" of the preparation of new TCM. To effectively promote the transformation of new TCM originating from the TCM clinical practice in medical institutions and establish an effective evaluation index system for the transformation of new TCM conforming to the characteristics of TCM, consensus experts adopted the literature research, questionnaire survey, Delphi method, etc. By focusing on the policy and technical evaluation of new TCM originating from the TCM clinical practice in medical institutions, a comprehensive evaluation from the dimensions of drug safety, efficacy, feasibility, and characteristic advantages was conducted, thus forming a comprehensive evaluation system with four primary indicators and 37 secondary indicators. The expert consensus reached aims to encourage medical institutions at all levels to continuously improve the high-quality research and development and transformation of new TCM originating from the TCM clinical practice in medical institutions and targeted at clinical needs, so as to provide a decision-making basis for the preparation, selection, cultivation, and transformation of new TCM for medical institutions, improve the development efficiency of new TCM, and precisely respond to the public medication needs.
Medicine, Chinese Traditional/standards* ; Humans ; Consensus ; Drugs, Chinese Herbal/therapeutic use* ; Surveys and Questionnaires

Beta-amylases (BAMs), key enzymes in starch hydrolysis, play an important role in plant growth, development, and resistance to abiotic stress. To mine the saline-alkali tolerance-related BAM genes in alfalfa (Medicago sativa L.), we identified MsBAM genes in the whole genome. The physicochemical properties, phylogeny, gene structures, conserved motifs, secondary structures, promoter cis-acting elements, chromosome localization, and gene replication relationships of BAM gene family members were analyzed. RNA-seq and quantitative real-time PCR (qRT-PCR) were employed to analyze the expression patterns of BAM family members under saline-alkali stress. The results showed that 54 BAM genes were identified in the genome, which were classified into 8 subgroups according to the phylogenetic tree. The members of the same subgroup had similar gene structures except that those of subgroups 1 and 7 had large differences. Conserved motif analysis showed that all MsBAM proteins had a typical glycohydrolysis domain. The chromosome localization analysis showed that MsBAM gene family members were unevenly distributed on 27 chromosomes. The duplication of gene segments led to the increase in BAM gene number in alfalfa. The promoters of BAM genes contained a large number of elements in response to plant hormones and stress. Transcriptome data and qRT-PCR results showed that the expression levels of most MsBAM genes were up-regulated in response to saline-alkali stress. Under the saline-alkali stress, the expression levels of 28 genes, including MsBAM6, were up-regulated on days 1 and 7, and those of 5 genes, including MsBAM9, were up-regulated by over 2 folds. In addition, under salt-alkali stress, BAM activity and soluble sugar content were significantly increased. These results indicate that BAM genes play a key role in alfalfa in response to saline-alkali stress, laying a foundation for further research in this field.
Medicago sativa/physiology* ; beta-Amylase/metabolism* ; Phylogeny ; Gene Expression Regulation, Plant ; Stress, Physiological/genetics* ; Multigene Family ; Alkalies ; Plant Proteins/genetics*

The connection between tendons and bones is called the tendon bone connection. With the continuous improvement of national sports awareness, excessive exercises and the related intensity are prone to damage the tendon bone connection. Tendon bone healing is a complex repair and healing process involving multiple factors, and good tendon bone healing is a prerequisite for its physiological function. The complexity of tendon bone structure also poses great challenges to the repair of tendon bone injuries. In recent years, researches have found that stem cells, growth factors, macrophages, and other factors are closely related to the healing process of tendon bone injuries, among which macrophages play an important role in the healing process. The authors reviewed relevant research literature in recent years and summarized the role of macrophages in tendon bone healing, in order to provide new ideas and directions for treatment strategies to promote tendon bone healing.
Humans ; Macrophages/metabolism* ; Wound Healing ; Animals ; Tendons/physiology* ; Bone and Bones/injuries* ; Tendon Injuries

Objective:To investigate the molecular characteristics and clinical heterogeneity of Usher syndrome（USH） -related gene variants in patients with hereditary hearing loss in southwest China, providing a basis for early diagnosis and clinical management. Methods:Thirteen patients from twelve families with hearing loss who attended the Affiliated Children's Hospital of Kunming Medical University between January 2017 and March 2021 were enrolled. All patients were identified as carrying USH-related gene variants through next-generation sequencing. Sanger sequencing was performed for all patients and their parents to validate the pathogenic variants. Comprehensive clinical evaluations, including medical history collection, otologic and ophthalmologic examinations, and vestibular function assessments, were conducted. Results:Among the 13 patients, 4 were diagnosed with USH type 1 and 2 with USH type 2. A total of 19 pathogenic or likely pathogenic variants were detected in USH-related genes, including MYO7A,CDH23,USH1C, and USH2A. The causative gene was MYO7A in 3 probands, CDH23 in 5, USH1C in 3, and USH2Ain 2. All patients exhibited an autosomal recessive inheritance pattern. Vestibular dysfunction was observed in 4 patients, and retinitis pigmentosa（RP） in 3 patients. Based on the genotype-phenotype correlation, 6 patients were initially diagnosed with USH, while 7 were classified as having non-syndromic hearing loss（NSHL）. Conclusion:This study revealed the clinical heterogeneity of USH-related gene variants in patients with hereditary deafness in southwest China. Although the clinical manifestations of USH are complex and there are overlapping characteristics between different subtypes, genetic testing provides an important basis for early diagnosis and precise clinical management. Especially for those with typical hearing loss, early genetic diagnosis can provide a window of time for early detection and intervention of retinitis pigmentosa.
Humans ; Usher Syndromes/genetics* ; Myosin VIIa ; Phenotype ; Male ; Female ; Myosins/genetics* ; Mutation ; Cadherins/genetics* ; Child ; Extracellular Matrix Proteins/genetics* ; Adolescent ; Pedigree ; High-Throughput Nucleotide Sequencing ; Cadherin Related Proteins ; Cytoskeletal Proteins ; Cell Cycle Proteins

Aim To explore the effect of neurite out-growth inhibitor A/neurite outgrowth inhibitor receptor(Nogo-A/NgR)pathway on psychedelic-reduced pre-pulse inhibition in mice.Methods Mice were injec-ted intraperitoneally with psilocybin,DOI to establish an animal model of prepulse inhibition(PPI)reduc-tion.The effects of psilocybin and DOI on PPI in mice after lateral ventricular injection of Nogo-A inhibitor NEP1-40 30 min in advance were evaluated.Finally,Rtn4r knockout mice were constructed to further verify the conclusion.Results The injection of NEP1-40(1 g·L-1,5 μL/mice,i.c.v)30 min in advance had no effect on PPI of mice.Under the conditions of 70 dB and 75 dB prepulse stimulation,NEP1-40 significantly up-regulated the PPI reduction induced by psilocybin.At the same time,NEP1-40 significantly up-regulated the DOI induced PPI reduction in mice at 70 dB and 80 dB prepulse stimulation.Compared with the two solvent groups,the PPI of Rtn4r-/-mice was not differ-ent from that of wild-type mice.Compared with the mice in Rtn4r-/-solvent group,the PPI of mice in Rtn4r-/-administration group showed a decreasing trend,but had no significant difference.Under the con-dition of 70 dB prepulse stimulation,there was a signif-icant difference between the Rtn4r-/-administration group and wild-type mice.Conclusion Nogo-A/NgR pathway is involved in the destruction of sensorimotor gating in mice by the psychedelic psilocybin or DOI.

Objective:To investigate effect of rapamycin on proliferation of human chronic lymphoblastic leukemia tumor cell MEC-1 in vitro.Methods:Human γδT cells were cultured and expanded in vitro.Human γδT cells percentage was detected by flow cytometry on the 10th day.γδT cells were treated with different concentrations of rapamycin(0,50,100 and 200 nmol/L).After 48 h of intervention,IL-17,IL-12,IFN-γ and TNF-α contents secreted by γδT cells were determined by ELISA,and AKT/mTOR protein expression was detected by Western blot.Supernatant of γδT was collected and co-cultured with MEC-1 indirectly.Co-cultured experi-mental was divided into DMSO group,rapamycin group,γδT group,rapamycin treated γδT group and control group.Survival rate of MEC-1 cells in each group was determined by CCK-8 after 48 h of culture.Results:IL-12,IFN-γ and TNF-α levels secreted by γδT cells were the highest when rapamycin was 100 nmol/L,which was significantly higher than other groups(P<0.05).When rapamycin was 50 nmol/L,IL-17 secretion level of γδT cells was the highest,and IL-17 level secreted by γδT cells gradually decreased with increase of rapamycin concentration.Survival rates of MEC-1 cells in supernatant of DMSO group,rapamycin group,rapamycin treated γδT cell group and γδT cell group were(93.48±2.52)%,(71.44±4.31)%,(83.93±1.54)%and(57.97±2.47)%,whose differences were statistically significant compared with control group(P<0.05).Compared with control group,mTOR protein level of γδT cells was decreased in all rapamycin treated groups.Protein expressions of upstream AKT,pAKT and downstream specific transcription factor 4EBP-1 were decreased with increase of rapamycin concentration(P<0.05).Conclusion:There is a dose-effect relationship between rapamycin concentration and IL-17,IL-12,IFN-γ,TNF-α secretions by γδT cells.Rapamycin and γδT cells can inhibit pro-liferation of MEC-1 cells,and 100 nmol/L rapamycin can enhance inhibitory effect of γδT cells on proliferation of MEC-1 cells.Rapa-mycin can weaken AKT pathway on γδT cells,and rapamycin combined with γδT cells plays a positive role in anti-tumor application of leukemia.

AIM To observe the protective effects and mechanism of Shuli Jiangzhuo Formula on cardiac function in a rat model of uremic cardiomyopathy(UCM).METHODS The successful UCM models established by 5/6 nephrectomy were randomly allocated into the model group,the valsartan group(8.33 mg/kg),and the low-dose,medium-dose and high-dose Shuli Jiangzhuo Formula groups(7.19,14.38,28.76 g/kg),in contrast to those of the sham operation group,followed by 8 weeks respective drug administration.Upon the completion of the pharmacological intervention,the rats had the left ventricular end-diastolic diameter(LVEDD),left ventricular end-systolic diameter(LVESD),ejection fraction(EF)and fractional shortening(FS)measured by echocardiography;the whole heart mass index(HMI)and left ventricular mass index(LVMI)detected;the renal function(serum creatinine,blood urea nitrogen)and the hemoglobin concentration detected;the mitochondrial morphology analyzed by observation of cardiomyocyte ultrastructure using transmission electron microscopy;the DNA damage in cardiomyocytes detected by TUNEL staining;the serum levels of IL-1β,IL-18 and BNP detected by ELISA;and the myocardial mRNA and protein expressions of NLRP3,Caspase-1 and IL-1β detected by RT-qPCR and Western blot.RESULTS Compared with the sham operated controls,the model group demonstrated significant elevation of serum creatinine and urea nitrogen(P＜0.01);decreased hemoglobin concentration(P＜0.01);disorganized myocardial collagen fiber architecture,and pronounced mitochondrial swelling with ultrastructural damage;decreased EF and FS(P＜0.05);increased LVEDD and LVESD(P＜0.01);increased HMI and LVMI(P＜0.01);increased levels of serum IL-1β,IL-18 and BNP(P＜0.01);increased cardiomyocyte pyroptosis(P＜0.01);and enhanced mRNA and protein expressions of NLRP3,Caspase-1 and IL-1 β(P＜0.01).Compared to model group controls,the high-dose Shuli Jiantuo Formula intervention exhibited decreased levels of serum creatinine and urea nitrogen(P＜0.01);increased hemoglobin concentration(P＜0.01);reduced DNA fragmentation,alleviated mitochondrial swelling and mitigated ultrastructural damage;reduced LVEDD and LVESD(P＜0.05,P＜0.01);decreased HMI and LVMI(P＜0.01);downregulated levels of serum IL-1β,IL-18 and BNP(P＜0.01);decreased cardiomyocyte pyroptosis(P＜0.01);and inhibited mRNA and protein expressions of NLRP3,Caspase-1 and IL-1β(P＜0.05,P＜0.01).CONCLUSION Shuli Jiangzhuo Formula demonstrates dual cardiorenal protective effects in UCM rats through suppression of the left ventricular hypertrophy progression and inhibition of the adverse ventricular remodeling processess.The therapeutic efficacy primarily stems from targeted suppression of NLRP3/Caspase-1 signaling pathway activation and substantial attenuation of cardiomyocyte pyroptosis cascade.