ObjectiveThis study aims to investigate the molecular mechanism by which Guizhi Fulingwan （GFW） inhibits ferroptosis in endometriosis （EMT） through the regulation of the hypoxia inducible factor-1α/heme oxygenase 1 （HIF-1α/HO-1） signaling pathway. MethodsMachine learning was employed to identify ferroptosis-related biomarkers associated with EMT. Network pharmacology was utilized to identify the active components of GFW and its potential therapeutic targets against EMT， including core targets. Functional enrichment analysis was conducted to explore the biological processes， molecular functions， cellular components， and signaling pathways associated with the potential targets. An EMT rat model was established via autologous transplantation. Thirty female Sprague-Dawley （SD） rats were randomly divided into five groups： sham-operated， model， positive control （dienogest at 0.2 mg·kg^-1）， low-dose GFW （2.5 g·kg^-1）， and high-dose GFW （5 g·kg^-1）. After modeling， the rats received their respective treatment by oral gavage for 28 consecutive days， while the sham and model groups received equal volumes of distilled water. Serum and ectopic endometrial tissues were collected. Hematoxylin and eosin （HE） staining was employed to evaluate morphological alterations in ectopic lesions. Quantitative real-time polymerase chain reaction （Real-time PCR） and Western blot were conducted to assess mRNA and protein expression of HIF-1α， HO-1， glutathione peroxidase 4 （GPX4）， spermidine/spermine N1-acetyltransferase （SAT1）， and prostaglandin-endoperoxide synthase 2 （PTGS2）. Tissue levels of malondialdehyde （MDA）， glutathione （GSH）， and ferrous iron （Fe²⁺） were quantified using commercial assay kits. Serum levels of interleukin-6 （IL-6） and transforming growth factor-β₁ （TGF-β₁） were measured via enzyme-linked immunosorbent assay （ELISA）. ResultsFive ferroptosis-related biomarkers in EMT were identified： ALOX12， CHAC1， SAT1， AST1， and HO-1. Network pharmacology analysis revealed 42 active components of GFW and 192 potential therapeutic target genes related to EMT treatment， with FOS， JUN， HO-1 identified as core targets. Functional enrichment analysis indicated that the potential targets were primarily involved in oxidative stress response and reactive oxygen species metabolism and were enriched in the HIF-1 signaling pathway. Compared to the sham-operated group， the model group exhibited significant increases in both mRNA and protein expression of HIF-1α， HO-1， and PTGS2， as well as elevated tissue levels of Fe²⁺ and MDA. Conversely， GSH levels and the expression of GPX4 and SAT1 were markedly reduced， and serum levels of IL-6 and TGF-β₁ levels were significantly higher （P<0.01）. Compared with the model group， all GFW-treated groups showed significant downregulation of HIF-1α and HO-1， reduced Fe²⁺ levels， and downregulated expression of MDA， PTGS2， IL-6， and TGF-β₁. Meanwhile， GSH， GPX4， and SAT1 expression levels were significantly increased （P<0.05， P<0.01）， effectively ameliorating iron overload and oxidative stress， thereby demonstrating therapeutic efficacy in EMT， with the high-dose GFW demonstrating the most pronounced therapeutic effects. ConclusionGFW exerts therapeutic effects on endometriosis by regulating the HIF-1α/HO-1 signaling pathway to rectify iron metabolism disorders and attenuate free iron-induced oxidative damage. It upregulates the antioxidative defense system to inhibit lipid peroxidation cascades and modulates inflammatory cytokine networks. These effects collectively disrupt the pathological interaction between ferroptosis and chronic inflammation， providing a novel theoretical foundation for the clinical application of GFW in EMT treatment.

Objective To investigate the multi-target mechanisms of quercetin in treating endometriosis(EMT)through integrative network pharmacology analysis.Methods Active targets of quercetin were collected from the TCMSP database,while EMT-related differentially expressed genes(DEGs)were identified through the Gene Expression Omnibus(GEO)dataset.A comparative analysis was conducted to pinpoint potential therapeutic targets of quercetin for EMT treatment.Functional enrichment analyses were employed to investigate the biological functions associated with these targets,and a protein-protein interaction(PPI)network was conducted to identify core targets.Molecular docking and dynamics simulations were performed to validate the binding characteristics between quercetin and the core targets.The top 2 target protein pairs,HSP90AB1 and AR,exhibiting the lowest binding energy,were selected for subsequent cellular experimental validation.Human EMT-immortalized ectopic endometrial epithelial cell line 12Z(n=6,independent replicates)was subjected,and CCK-8 assay was used to determine ehe effects of quercetin on cell viability and proliferation,and the half-maximal inhibitory concentration(IC50)was calculated at 48 h after treatment.Then the 12Z cells were treated with quercetin at a concentration gradient of 0,30,60 and 90 μmol/L,the migration and invasion abilities were assessed with cell scratch and cell invasion assays.Western blotting was conducted to detect the changes in the expression of HSP90AB1 and AR proteins after different doses of treatment.Results There were 49 potential EMT-related therapeutic targets and 10 core targets identified.Functional enrichment analyses revealed that these targets were significant enriched in inflammation-related signaling pathways,including AGE-RAGE,ErbB and TNF;immune-related pathways,such as Th17 cell differentiation,T/B cell receptor signaling;angiogenesis-related pathways like VEGF;and hormonal regulatory pathways involving estrogen and GnRH.Molecular docking demonstrated that quercetin exhibited favorable binding activity(binding energy<-5 kcal/mol)with all core target proteins,with particularly strong binding energies(<-7 kcal/mol)observed for AR,EGFR,FOS,ERBB2,and HSP90AB1.Molecular dynamics simulations revealed that quercetin forms sustained hydrogen bond interactions with AR and HSP90AB1,facilitating the formation of stable complexes.CCK-8 assay,cell scratch assay,and transwell invasion assay indicated that quercetin inhibited the proliferative activity,and migrative and invasive abilities of 12Z cells in a concentration-dependent manner,with more pronounced inhibitory effects observed at 60 and 90 μmol/L quercetin(P<0.001);Western blotting revealed that treatment of 12Z cells with varying quercetin concentrations for 48 h up-regulated the expression of HSP90AB1 and AR,with the most significant increase observed at 90 μmol/L quercetin(HSP90AB1,P<0.05;AR,P<0.001).The restored expression levels of HSP90AB1 and AR showed positive correlations with the proliferative activity,migrative and invasive abilities of ectopic endometrial cells.Conclusion Quercetin effectively addresses endometriosis through multiple molecular targets and signaling pathways,and stabilization of the HSP90AB1/AR complex and subsequent protein upregulation represents a key therapeutic mechanism.

Objective:To explore the impact of different family management patterns on the family resilience of children with brain tumors.Methods:A total of 210 parents of children with postoperative brain tumors in 2 tertiary Grade A pediatric hospitals were investigated by the general information questionnaire, family management measure (FaMM) and family resilience rating scale.SPSS 24.0 was used for cluster analysis.Results:(1) Among the scores of FaMM, child identity, condition management ability and parental mutuality were significantly positively correlated with family resilience ( r=0.312, r=0.470, r=0.391, all P<0.05), while view of condition impact, condition management difficulty and condition management effort were negatively correlated with family resilience ( r=-0.346, r=-0.177, r=-0.348, all P<0.05). (2) Family management patterns could be divided into four categories: burden managing (22.9%), effective managing (24.8%), poor managing (28.6%) and tacit managing (23.8%). (3) Family resilience scores of the four patterns were (197.21±20.08), (205.92±14.25), (181.47±18.13) and (198.06±17.08), and their differences were statistically significant ( F=19.498, P<0.01). Conclusion:The family resilience level of children with brain tumor is associated with different family management patterns.Therefore, effective strategies according to different family characteristics should be developed to improve family resilience level.

In this study, the distribution of five Alzheimer's disease (AD)-related single nucleotide polymorphisms (SNPs) in the Han population was examined in combination with the evaluation of clinical cognition and brain pathological analysis. The associations among SNPs, clinical daily cognitive states, and postmortem neuropathological changes were analyzed in 110 human brains from the Chinese Academy of Medical Sciences/Peking Union Medical College (CAMS/PUMC) Human Brain Bank. APOE ε4 (OR = 4.482, P = 0.004), the RS2305421 GG genotype (adjusted OR = 4.397, P = 0.015), and the RS10498633 GT genotype (adjusted OR = 2.375, P = 0.028) were associated with a higher score on the ABC (Aβ plaque score, Braak NFT stage, and CERAD neuritic plaque score) dementia scale. These results advance our understanding of the pathogenesis of AD, the relationship between pathological diagnosis and clinical diagnosis, and the SNPs in the Han population for future research.
ADAM10 Protein ; genetics ; Adult ; Aged ; Aged, 80 and over ; Alzheimer Disease ; genetics ; pathology ; Amyloid Precursor Protein Secretases ; genetics ; Antiporters ; genetics ; Apolipoprotein E4 ; genetics ; Asian Continental Ancestry Group ; genetics ; Brain ; pathology ; Cognitive Dysfunction ; genetics ; pathology ; Female ; Genetic Predisposition to Disease ; Humans ; Male ; Membrane Proteins ; genetics ; Middle Aged ; Polymorphism, Single Nucleotide

Brain ; pathology ; China ; Humans ; Organ Preservation ; standards ; Tissue Banks ; ethics ; standards

OBJECTIVETo compare the clinical efficacy difference between encircling needling combined with physical factor therapy and simple physical factor therapy for severe pressure sore, and to explore the optimal method for severe pressure sores.

METHODSThirty-four patients with IV-grade pressure sore were randomly divided into an observation group and a control group, 17 cases in each one. Patients in the control group were treated with conventional nursing, ultrasonic wave and short-wave ultraviolet therapy; additionally, the encircling needling was applied in the observation group. All the treatment was given once a day, 5 times a week, and 4-week treatment constituted one session. Totally, two sessions of treatment were performed. Three indices, including the area of pressure sore, 24-h volume of exudates and wound-bed tissue type, were compared between the two groups before and after treatment; the clinical efficacy was evaluated in the two groups.

RESULTSAfter treatment of one session and two sessions, the area of pressure sore, 24-h volume of exudates and wound-bed tissue type were significantly reduced in the two groups (P < 0.01, P < 0.05), which was more obvious in the observation group (all P < 0.05). The total effective rate in the observation group was 76.5% (13/17) after 1 session and 94.1% (16/17) after 2 sessions, which were superior to 35.3% (6/17) after 1 session and 64.7% (11/17) after 2 sessions in the control group (both P < 0.05).

CONCLUSIONEncircling needling combined with physical factor therapy can obviously reduce the pressure sore area and 24-h volume of exudates and improve wound-bed tissue type, which is superior to simple physical factor therapy.

Acupuncture Therapy ; Adult ; Aged ; Combined Modality Therapy ; Female ; Humans ; Male ; Middle Aged ; Pressure Ulcer ; therapy ; Short-Wave Therapy ; Ultrasonic Therapy ; Ultrasonic Waves

Objective To study the efficacy of intra-tumoral injection of different concentrations of ethanol for nude mice with implanted pancreatic cancer and provide evidence for choosing appropriate concentration of ethanol for clinical treatment of pancreatic cancer.Methods A subcutaneous xenograft mouse model of human pancreatic cancer SW1990 was established.Forty-eight nude mice with similar tumor size were randomly divided into 20％,40％,60％,80％,95％ ethanol injection groups and saline injection group.The longest (a) and the shortest diameters (b) of tumor of nude mice were measured.Tumor volume (TV),relative tumor volume (RTV) and the relative rate of tumor proliferation (T/C％) were calculated.Eight days later the nude mice were sacrificed.The tumor tissue was harvested for pathologic examinations.Results RTV in 20％ ethanol injection group was similar that of saline injection group (P =0.212).RTV in 40％,60％,80％ and 95％ ethanol injection groups were significantly lower than that in saline injection group (P ＜ 0.01).RTV was less than 1 and T/C％ was less than 30％ in 60％,80％ and 95％ ethanol injection groups.The values of RTV and T/C％ decreased with the increase of ethanol concentration.RTV in 80％ and 95％ ethanol injection groups were significantly lower than that of 60％ ethanol injection group (P =0.003 and P =0.009).RTV was similar in 80％ and 95％ ethanol injection groups (P =0.819).The pathologic examinations showed no tumor necrosis in saline injection group,while small amounts of necrosis in implanted pancreatic cancer was observed in 20％ and 40％ ethanol injection groups,while a large area of coagulation necrosis could be found in 60％,80％ and 95％ ethanol injection groups.Conclusions Intra-tumoral injection of 80％ ethanol is feasible therapy method for nude mice with human pancreatic cancer xenografts.

To evaluate the effectiveness of rabies vaccination, we developed the SPA-ELISA method to detect the antibodies against rabies virus (RV) using the main antigenic determinant of nucleoprotein (RV N1) as antigen. The complete Nucleoprotein (N) gene and the partial N1 gene (1 000-1 353 bp) of RV Flury LEP strain were amplified using RT-PCR and PCR approaches. The two fragments were inserted into pGEX-6P-1 respectively. Then we transformed the recombinant plasmids into Escherichia coli BL21(DE3) strain and expressed them by adding 1 mmol/L of IPTG (isopropyl-beta-D-thiogalactopyranoside). SDS-PAGE analysis showed that both of the two recombinant proteins were presented as inclusion bodies. Compared with the complete nucleoprotein, the partial protein (RV N1) was expressed at a much higher level in E. coli BL21(DE3). The antigenic specificity of the partial N1 protein was confirmed by Western blotting. By coating the plates with purified RV N1 as an antigen, an SPA-ELISA method for the detection of the antibodies against RV was established. By optimizing this method, the optimal concentration of RV N1 coating the ELISA plate was 2 mg/L. The optimal concentration of serum samples and SPA-HRP was 1:100 and 1:4 000 respectively. Compared with a commercially available ELISA kit coating RV as antigen, the coincidence rate of SPA-ELISA was 94.1%. Our results show that the developed SPA-ELISA based on the RV N1 was useful for the detection of the antibodies against RV in the sera of dogs.
Animals ; Antibodies, Viral ; analysis ; immunology ; Dogs ; Enzyme-Linked Immunosorbent Assay ; methods ; Epitopes ; immunology ; Escherichia coli ; genetics ; Genetic Vectors ; genetics ; Nucleocapsid Proteins ; immunology ; Rabies virus ; immunology ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Staphylococcal Protein A

Objective To investigate if improving or slowing the progression of glucose intolerance might be linked with the reduction of cardiovascular disease(CVD)events and mortality in a Da Qing population with prediabetes.Methods In 1986,577 subjects with impaired glucose tolerance in 33 clinics in Daqing city were randomly assigned to either the control group or one of three lifestyle intervention groups(diet,exercise,or diet plus exercise)to receive a 6 year lifestyle intervention.All the participants were followed for 14 years(1993-2006)after completion of the 6 year active interventions to assess the long-term effect of the interventions.In this post-hoc analysis,the participants were stratified into four subgroups(quartiles)based on their 2 h plasma glucose(2hPG)level after glucose loading at the end of the active intervention,in order to analyze the impact of plasma glucose level on CVD events and mortality.Results During the 20-year follow-up,there were a total of 142 deaths(68 of which were attributed to CVD)and 211 first CVD events(145 strokes and 66 myocardial infarctions).From the highest to the lowest levels of 2hPG in the 4 quartiles,the all-cause mortality(17.8,12.7,10.9,and 9.7/1 000 personyears),CVD mortality(9.1,5.9,6.1,and 4.9/1 1300 person-years)and the incidence of first CVD events(30.4.24.0,18.8,and 19.7/1 000 person-years)showed a clear trend of decline.In multivariate analyses,controlled for age,sex,body mass index,smoking habit,blood pressure,and intervention methods at baseline,the results showed that the 5 mmol/L elevation of 2hPG level after glucose loading in 1992 significantly increased the all-cause mortality(HR 1.335.P=0.005),the incidences offirst CVD events(HR 1.227,P=0.012)and stroke(HR 1.213,P=0.026).Conclusion In pre-diabetes population.if the lifestyle intenrentions are substantially efficacious in improving glucose intolerance,the CVD risk and mortality will be reduced.

Objective To investigate the effect of mesenchymal stem cells (MSCs) on subcutaneous xenograft tumors in mice with Lewis lung cancer. Methods MSCs isolated from bone marrow of C57BL/6 mice were made into single cell suspension and were cultured in vitro. The cells of the 4th to 5th passage were used for the subsequent experiments. Fifty six C57BL/6 mice were inoculated subcutaneously with Lewis lung cancer cells, and were grouped into Group D0 (MSCs were given simultaneously with inoculation)and Group D10(MSCs were given 10 d after inoculation). Group D0 included three subgroups (n=8): Group 1 with inoculation of tumor cells, Group 2 with inoculation of tumor cells and MSCs, and Group 3 with inoculation of tumor cells and tail intravenous injection of MSCs. Group D10 included four groups: Group 4 with inoculation of tumor cells and injection of MSCs in tumors, Group 5 with equivalent PBS (the control of Group 4), Group 6 with inoculation of tumor cells and tail intravenous injection of MSCs, and Group 7 with equivalent PBS (the control of Group 6). The time of tumor formation and the volume of tumor were observed and compared among the groups. ResultsIn Group D0, earlier onset of tumor development was observed in Group 2 as compared to Group 1 and Group 3 (P<0.05), while there was no significant difference on the volume of tumor in the three groups (P>0.05). In Group D10, the volume of tumors were larger in Group 4 compared to the control (P<0.05), while there was no significant difference on the volume of tumors between Group 6 and the control (P>0.05). Conclusion Inoculating mixture of MSCs and Lewis lung cancer cells accelerates tumor formation,and injection of MSCs in tumors stimulates the growth of tumors.