BACKGROUND:Ferroptosis is a mode of programmed cell death distinct from apoptosis,necrosis,and other novel cellular deaths,which occurs mainly due to accumulated lipid peroxidation.Ferroptosis has been shown to be involved in the pathological process following subarachnoid hemorrhage.Baicalein,serving as an adept sequestered of iron,evinces its prowess by quelling lipid peroxidative cascades.Nonetheless,the enigma lingers as to whether baicalein possesses the capacity to ameliorate neuronal ferroptosis,elicited in the wake of early brain injury after subarachnoid hemorrhage. OBJECTIVE:To investigate the effect and mechanism of baicalein on neuronal ferroptosis after subarachnoid hemorrhage. METHODS:Primary neuronal cells were extracted from C57BL/6L fetal mice at 16-17 days of gestation.Hemoglobin was used to stimulate primary neuronal cells to simulate an in vitro subarachnoid hemorrhage model.The viability of primary neuronal cells treated with baicalein at concentrations of 5,15,25,50,and 100 μmol/L for 24 hours was detected by CCK-8 assay to determine the optimal concentration of baicalein.Primary neuronal cells were divided into control group,hemoglobin group,and hemoglobin+baicalein group.The levels of reactive oxygen species and malondialdehyde in cells were detected by kits.The mRNA expressions of ferroptosis-related markers PTGS2,SLC7A11,and glutathione peroxidase 4 were detected by RT-PCR.The primary neuronal cells were further divided into control group,SLC7A11 inhibitor Erastin group,hemoglobin group,hemoglobin+baicalein group,and hemoglobin+baicalein+Erastin group.The expression of the ferroptosis related markers SLC7A11 and glutathione peroxidase 4 was detected by western blot assay. RESULTS AND CONCLUSION:(1)Baicalein(25 μmol/L)was selected as the following experimental concentration.(2)Compared with the hemoglobin group,the level of malondialdehyde and the level of reactive oxygen species were significantly decreased(P<0.05)in the hemoglobin+baicalein group.(3)Compared with the hemoglobin group,the mRNA expression of PTGS2 significantly decreased,and the mRNA expression of SLC7A11 and glutathione peroxidase 4 significantly increased(P<0.000 1)in the hemoglobin+baicalein group.(4)SLC7A11 inhibitor Erastin could reverse the baicalin-improved ferroptosis effect to a certain extent(P<0.05).(5)The results showed that baicalein could alleviate the ferroptosis of neuronal cells after subarachnoid hemorrhage through the SLC7A11/GPX4 pathway.

Post-traumatic stress disorder (PTSD) is a relatively common but complex mental illness with a range of diverse risk factors. Typical symptoms include the re-experience or avoidance of traumatic events, cognitive impairment, and hypervigilance. While the exact pathogenesis of PTSD is unclear, many studies indicate that epigenetic regulation plays a key role in its development. Specifically, numerous studies have indicated that the levels of histone acetylation and methylation, DNA methylation, and noncoding RNA are altered in PTSD patients. Further to this, natural products have been found to achieve epigenetic regulation of PTSD by regulating the expression of epigenetic enzymes, long noncoding RNA (lncRNA), and miRNA, thereby playing a role in improving PTSD symptoms. To date, however, no epigenetic regulation related drugs have been used in the treatment of PTSD. Furthermore, while natural products that can epigenetically regulate PTSD have received increasing levels of attention, there have not yet been any systematic reports on the topic. Here, we summarized the roles and mechanisms of natural products in the epigenetic regulation of PTSD, providing a novel and unique perspective that will help to guide the development and application of new PTSD treatments.

ObjectiveThis paper used the AGREE Ⅱ guideline evaluation tool to evaluate the quality of 14 clinical guidelines for acute myocardial infarction，aiming to provide reference for the formulation and improvement of the guidelines. MethodsClinical guidelines and expert consensus related to acute myocardial infarction were searched by web search. The search period ranges from January 1，2019 to November 1，2024 in CNKI，VIP，Wanfang Data，SinoMed，Web of Science，OVID， the International Guidelines Collaboration Network （GIN），the UK National Institute for Health and Clinical Excellence （NICE），Yimaitong， and other platforms. Three researchers independently screened the literature and used AGREE Ⅱ to score the screening results. After ensuring that the researchers have a consistent understanding of each guideline，the quality of the guidelines was evaluated. After that，the ratings were analyzed by layer according to the issuing agency，category，method of formulation，and funding situation and compared longitudinally by rating time. The clinical guidelines and expert consensus were compared in terms of content and evidence. ResultsA total of 14 guidelines and consensus were included. The results of AGREE Ⅱ in the six areas in descending order were scope and purpose （62.82%±10.43%），rigor （62.40%±12.77%），editorial independence （62.11%±22.26%），participants （61.42%±11.65%），clarity of expression （59.98%±9.62%），and application （52.94%±16.90%） . Eleven of the guidelines were at level B， and three were at level A. In the stratified analysis，the score of the guideline formulated by the Chinese Medical Doctor Association was lower. There was little difference between the scores of Chinese/Western and Western medicine guidelines. The average score of the guidelines was higher than the consensus. Funded guidelines and consensus scores were higher. In the longitudinal comparison，the highest number of guidelines were developed in 2020 and 2021，while those developed in 2023 scored the highest. In the differential comparison analysis，the content of the guidelines was more comprehensive， and the evidence level was higher，while the content of the consensus was more novel， and the evidence was less. ConclusionThe AGREE Ⅱ score of the clinical guidelines for acute myocardial infarction is generally moderate，and there is room for improvement in terms of applicability. At the same time，the content quality of expert consensus should be improved，and more efforts should be made to develop and apply Chinese medicine guidelines for complications such as heart failure and microcirculatory obstruction after acute myocardial infarction.

Objective:To analyze the role of interleukin-33(IL-33)in the occurrence and development of glioma and its related mechanism by bioinformatics technology,and to validate it through histopathological experiments,and to discuss the possibility of IL-33 as an auxiliary marker for the diagnosis and treatment of brain glioma.Methods:The glioblastoma multiforme/lower grade glioma(GBMLGG)case data were downloaded from the UCSC XENA database,including data of 689 glioma samples,5 paracancerous samples,and 1 152 normal brain tissue samples;Mann-Whitney U test was used to analyze the difference in the expression of IL-33 mRNA between the GBMLGG samples and the normal brain tissues;according to the expression level of IL-33 in GBMLGG tissue,the tumor samples were divided into IL-33 low expression group and IL-33 high expression group;the Human Protein Atlas(HPA)was used to validate the difference in the protein expression of IL-33 in the GBMLGG samples;the R language DESeq2(v.1.36.0)package was used to screen the differentially expressed genes(DEGs)in the GBMLGG tumor case samples;Gene Ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway enrichment analysis were used to perform pathway analysis on the DEGs;Gene Set Enrichment Analysis(GSEA)was used to discuss the pathways significantly enriched by IL-33 in the GBMLGG tissues;GSVA package was used to analyze the immune infiltration in the GBMLGG samples;survival package and survminer package were used to analyze the effect of IL-33 expression level on the survival of the patients in different clinical subgroups of GBMLGG;univariate and multivariate Cox proportional hazards regression models were used to analyze the relationship between IL-33 expression and the clinicopathological characteristics of the GBMLGG patients;the GBMLGG and control tissue samples were collected;immunohistochemical staining was used to detect the expression levels of IL-33 and its receptor suppression of tumorigenicity 2(ST2)in the GBMLGG and normal brain tissue samples.Results:The expression levels of IL-33 mRNA and protein in the GBMLGG tissues were significantly increased compared with those in normal brain tissues;there were 634 DEGs in total between the IL-33 low and high expression groups,including 283 up-regulated DEGs and 351 down-regulated DEGs;the GO functional enrichment analysis and KEGG signaling pathway enrichment analysis results showed that the DEGs were associated with biological behaviors such as activation of the classical pathway of complement,immunoglobulin complex formation,and mediated immunoglobulin receptor binding;in the course of GBMLGG development,high expression of IL-33 could degrade valine,leucine,and isoleucine,induce limonene and pinene degradation,promote propanoate metabolism,and simultaneously activate the Leishmania infection pathway,NOD-like receptor signaling pathway,and allograft rejection pathway;the infiltration levels of dendritic cell(DC)and mast cell in the IL-33 high expression group were higher than those in IL-33 low expression group;the infiltration levels of eosinophil,helper T cell,and central memory T cell(Tcm)were lower than those in IL-33 low expression group;the expression level of IL-33 was positively correlated with the infiltration of γδT cell(Tgd),helperT cell,macrophage,eosinophil,Tcm,and effector memory T cell(Tem)(P＜0.05);it was negatively correlated with the infiltration levels of DC,natural killer cell(NK),CD8+T cell,and CD56bright NK cell(P＜0.05).There were no significant differences in the overall survival(OS),disease-specific survival(DSS),and disease-free interval(DFI)of the GBMLGG patients between IL-33 high expression group and IL-33 low expression group(P＞0.05);the clinical subgroup analysis results showed that the expression level of IL-33 in oligodendrocytoma tissues was lower than those in astrocytoma and oligoastrocytoma tissues,and the expression level of IL-33 in glioblastoma tissues was higher than that in oligodendroglioma tissues.World Health Organization(WHO)stage and age were risk factors affecting the prognosis of the GBMLGG patients,and IDH mutation and primary treatment effect were protective factors affecting the prognosis;The immunohistochemical staining results showed that compared with normal brain tissues,the expression levels of IL-33 and its receptor ST2 proteins in the malignant glioma tissues were significantly increased(P＜0.05),and their expression levels were positively correlated in both normal brain tissues and malignant glioma tissues(P＜0.05).Conclusion:The expression level of IL-33 in the glioma tissue is significantly increased,and high expression of IL-33 may be a potential factor for poor prognosis in the glioma patients.

OBJECTIVE: A new three-dimensional(3D) classification of posterior cruciate ligament (PCL) tibial avulsion fracture based on computed tomography(CT) features was established and the significance in clinical treatment was explored in this study. METHODS: From May 2013 to November 2023, 43 cases of PCL tibial avulsion fracture in the Second Hospital of Jilin University were analyzed retrospectively, including 29 males and 14 females, aged (34.3±8.5) years. According to traditional Meyers and McKeever classification, 3 cases were typeⅠ;2 cases of typeⅡ;38 cases were type Ⅲ. Based on the characteristics of CT images, 43 patients were given specific treatment strategies and followed up to evaluate the curative effect. According to the degree of fracture displacement, involved range and the integrity of fracture block demonstrated by CT images, the new three-dimensional classification of PCL avulsion fracture was established. Kappa coefficient was used for consistency test. RESULTS: A new 3D classification of PCL tibial avulsion fracture was established. TypeⅠwas the non-displaced fracture (displacement degree ≤3 mm), in which typeⅠa was the avulsion range limited in the posterior intercondylar fossa, and Ib was the avulsion range beyond the posterior intercondylar fossa. TypeⅡrepresented the displaced fracture in the posterior intercondylar fossa (avulsion limited to the posterior intercondylar fossa and fracture displacement>3 mm), in which typeⅡa represented a slight displacement with a intact broken block and the posterior elevation of the avulsion (hinge mechanism), typeⅡb represented the complete separation of fracture ends with a intact fracture block, and typeⅡc was the comminuted fracture. Type Ⅲ was the displaced fracture beyond the posterior intercondylar fossa (avulsion involving the articular surface of the tibial plateau or the intercondylar ridge and the degree of displacement > 3 mm), among which type Ⅲa was the simple fracture with intact broken block, type Ⅲb represented the comminuted fracture, and type Ⅲc was the complex fracture with tibial plateau fracture. According to this new 3D classification, 43 patients were classified as type Ia in 2 cases and typeⅠb in 1 case;typeⅡa in 2 cases, typeⅡb in 15 cases and typeⅡc in 7 cases;type Ⅲa in 2 cases, type Ⅲb in 5 cases and type Ⅲc in 9 cases. All the 43 cases in this study achieved bone union. At the last follow-up, according to the hospital for special surgery knee score(HSS)evaluation system for the knee joint function, 27 cases were excellent, 11 cases were good, 5 cases were fair. The average Kappa value of inter-observer reliability in the first stage was 0.793, and the second stage was 0.855. The average Kappa value of the whole stage was 0.839, indicating high level of consistency. The average Kappa value of intra-observer reliability was 0.893, indicating high level of consistency. CONCLUSION The 3D classification of PCL tibial avulsion fracture is intuitive, demonstrating a high level of reliability. It has a certain guiding significance for the selection of clinical treatment methods, and it is suggested to be promoted and applied as a new classification system in clinical practice.
Humans ; Male ; Female ; Posterior Cruciate Ligament/surgery* ; Adult ; Tibial Fractures/classification* ; Tomography, X-Ray Computed ; Middle Aged ; Retrospective Studies ; Fractures, Avulsion/classification* ; Imaging, Three-Dimensional ; Young Adult

This study was aimed at investigating the effectiveness of various host nucleic acid removal and non-specific amplifica-tion techniques in animal tissue samples,to increase the accuracy of pathogen identification in tissue samples.Simulated samples were prepared with a mixture of mouse lung tissue homogenates and Klebsiella pneumoniae fluids,and processed with six host nucleic acid removal kits and three non-specific amplification techniques.The effectiveness of each method in removing host DNA and enriching nucleic acids of pathogenic microorganisms was evaluated through real-time fluorescence quantitative PCR and high-throughput se-quencing.For host nucleic acid removal techniques,the method of selective cleavage and quantitative degradation of host DNA(Com-plete5 kit)effectively decreased the host nucleic acid content in tissue samples and increased the relative abundance of pathogen nucleic acids.In contrast,the magnetic bead method for host DNA removal(Next microbiome DNA enrichment Kit kit)was less effec-tive.At lower pathogen concentrations(77 CFU/mL),the Vazyme kit was more effective than the other kits in removing host nucleic acids.Non-specific amplification techniques(MALBAC whole genome amplification,MDA isothermal amplification,and random primer amplification)were not applicable to tissue samples and were not effective in increasing the relative abundance of pathogen nucleic acids.Selective lysis and quantitative degradation of host DNA were suitable for processing tissue samples with high host back-ground and low pathogenic microorganism levels,whereas non-specific amplification methods were not applicable to tissue samples for pre-processing of macro-genome high-throughput sequencing.

Objective To investigate the relationship between preoperative serum high mobility group protein 1(HMGB1),macrophage inflammatory protein-1α(MIP-1α),osteopontin(OPN)and the prognosis of patients with basal ganglia intracerebral hemorrhage after neuroendoscopic surgery.Methods From March 2022 to March 2024,98 patients with cerebral hemorrhage in basal ganglia treated by neuroendoscopic removal were selected.Serum HMGB1,MIP-1 a and OPN were detected before operation,and they were followed up for 1 month after operation.Cox regression analysis was used to analyze the risk factors affecting the prognosis of patients with cerebral hemorrhage in basal ganglia.Kaplan-Meier survival curve and Log-rank test were used to analyze the survival rate of patients with cerebral hemorrhage in basal ganglia.Results Among the 98 patients with cerebral hemorrhage in basal ganglia,21 patients died within 1 month after neuroendoscopic removal,and the mortality rate was 21.43％.The proportion of patients with midline shift ≥ 10 mm(76.19％)and hematoma volume[(51.18±7.62)ml]in the death group were higher than those in the survival group[14.29％and(31.93±5.66)ml],and the hematoma clearance rate[(78.13±5.86)％]was lower than that in the survival group[(90.58±6.29)％](P＜0.05).The preoperative levels of HMGB1,MIP-1 α and OPN in the death group were(12.44±2.26)ng/ml,(417.25±134.12)pg/ml and(12.32±3.19)ng/ml,respectively.The survival groups were(7.52±2.37)ng/ml,(200.35±31.36)pg/ml and(6.29±2.37)ng/ml,respectively.There was a statistically significant difference between the two groups(P＜0.05).Cox regression analysis showed that preoperative high HMGB1(HR=1.629,95％CI:1.274-2.083),high MIP-1α(HR=2.875,95％CI:1.384-5.972),high OPN(HR=1.429,95％CI:1.073-1.093)were risk factors for death within 1 month after neuroendoscopic removal of basal ganglia intracerebral hemorrhage(P＜0.05).Survival curve analysis showed that the survival rate of high HMGB1,high MIP-1α and high OPN at 1 month after operation was significantly lower than that of low HMGB1,low MIP-1α and low OPN(Long-rank x2=7.539,6.028,8.220,P＜0.05).Conclusion The prognosis of patients with basal ganglia intracerebral hemorrhage with high HMGB1,high MIP-1α and high OPN before neuroendoscopic surgery was poor.Preoperative HMGB1,MIP-1α and OPN may be used as indicators for postoperative evaluation of neuroendoscopic removal in patients with basal ganglia intracerebral hemorrhage.

Objective To investigate the protective effects and mechanism of Shuilian Fengkui Formula(SFF)of Yao medicine on sequelae pelvic inflammatory disease(SPID)in rats.Methods The SPID rat model was established by mechanical injury coupled with bacterial infection,and then the rats were randomly divided into:the SPID group,the positive drug(Fukeqianjin tablets)group,SFF groups at low,medium and high doses,and the sham-operation group,with 8 rats in each group;SPID group,SFF groups,TLR4 agonist CRX-527(CRX-527)group,SFF plus CRX-527 group,with 8 rats in each group.Corresponding drug intervention was given for 21 consecutive days.The indexes of uterus,spleen,thymus and other organs were calculated.The histopathological changes in rat uterus were observed by HE staining.The levels of interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),immunoglobulin G(IgG),immunoglobulin M(IgM)in serum were detected by ELISA;The protein expression levels of Toll receptor 4(TLR4),myeloid differentiation factor(MyD88),nuclear factor kappa B p65(NF-κB p65),and phosphorylated nuclear factor kappa B p65(p-NF-κB p65)in rat uterine tissues were detected by Western blot.Results Compared with the sham operation group,the uterine index of SPID group significantly increased(P<0.01),the spleen index and thymus index significantly decreased(P<0.01),the pathological damage of uterine tissue was serious,the contents of IL-1β,IL-6,TNF-α in serum significantly increased(P<0.01),the contents of IgG and IgM significantly decreased(P<0.01),the TLR4,MyD88 protein and the ratio of p-NF-κB p65/NF-κB p65 protein in uterine tissue significantly increased(P<0.01).Compared with the SPID group,the uterine index of the positive drug group and the SFF groups significantly decreased(P<0.05),the spleen index and thymus index significantly increased(P<0.05),the pathological damage of uterine tissue significantly improved,the contents of IL-1β,IL-6 and TNF-α in serum were significantly lowered(P<0.05),the contents of IgG and IgM significantly increased(P<0.05),and the TLR4,MyD88 protein and the ratio of p-NF-κB p65/NF-κB p65 protein in uterine tissue significantly downregulated(P<0.01).However,CRX-527 significantly reversed the effects of SFF on SPID in rats.Conclusion SFF could improve SPID in rats by impeding the inflammatory response,and its mechanism might be attributed to inhibition of TLR4/MyD88/NF-κB signaling pathway.

AIM To study the chemical constituents from Stelmatocrypton khasianum.(Benth.)Baill.and their in vitro anti-inflammatory activity and cytotoxic activity.METHODS Separation and purification were performed using thin layer chromatography,silica gel,semi-preparative HPLC and Sephadex LH-20,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The in vitro anti-inflammatory activity was evaluated by RAW264.7 model,and the cytotoxic activity was determined by CCK-8 method.RESULTS Fifteen compounds were isolated and identified as 2α,3β,19α,23-tetrahydroxy-urs-12-en-28-oic acid-3-O-β-D-glucopyranosyl-28-O-β-D-glucopyranosyl ester(1),dalzienoside(2),benzyl-(6-O-α-L-rhanmopyranosyl)O-β-D-glucopyranoside(3),corchorusoside C(4),cyclo(Ala-Tyr)(5),thymidine(6),(4S,5S)-5-hydroxy-4-hexanolide(7),2-methylpyridin-3-ol(8),butyl isobutyl phthalate(9),bis-(2-ethylhexyl)terephthalate(10),p-hydroxybenzaldehyde(11),vanilic acid(12),salicylic acid(13),isovanilic acid(14),3-hydroxy-p-anisaldehyde(15).The IC50 values of compounds 1,2 and 4 for NO were(27.69±5.51),(25.82±3.58)and(23.35±7.09)μmol/L,respectively.The IC50 value of compound 1 on MCF-7 cells was(18.15±6.45)μmol/L.The IC50 values of compound 4 on MCF-7 and HCCC-9810 cells were(19.43±2.66)and(21.76±5.81)μmol/L,respectively.CONCLUSION Compounds 2-11 are isolated from S.khasianum for the first time.Compounds 1,2 and 4 exhibit good in vitro anti-inflammatory activity,and 1,4 have cytotoxic activity.

Objective:To observe the effect of robot-assisted gait training (RAGT) supplemented with intermittent theta burst stimulation (iTBS) of the cerebellum in restoring lower limb function after anterior cruciate ligament reconstruction (ACLR).Methods:Eighty ACLR patients were randomly divided into a control group, a magnetic stimulation group, a robot group and a combined group, each with 20 members. The robot and magnetic stimulation groups underwent RAGT and cerebellar iTBS before conventional training, while the combined group received iTBS followed by RAGT and then conventional training. The treatments were administered once a day, three days per week for four weeks. Before and after the intervention, the peak torque ratio of the knee flexors and extensors (H/Q), peak torque of the knee extensors (PT), and knee repositioning angle difference were measured. Knee function and balance (using the Berg Balance Scale (BBS)) were also assessed.Results:The combined group demonstrated significantly better quadriceps PT and H/Q% than the other 3 groups. Knee repositioning angle difference improved significantly in all of the groups after the treatment, with the combined group showing the smallest difference (5.00±1.21)°, significantly better than the other three groups. Lysholm and BBS scores had also improved significantly in all of the groups, with the combined group′s improvements again significantly better than those of the other groups.Conclusion:Intermittent theta burst stimulation of the cerebellum combined with robot-assisted gait training can significantly improve knee function and balance after ACLR.